C-terminal modified oxamyl dipeptides as inhibitors of the ICE-ced-3 family of cysteine proteases

ABSTRACT

This invention is directed to novel oxamyl dipeptide ICE/ced-3 family inhibitor compounds. The invention is also directed to pharmaceutical compositions containing these compounds, as well as to the use of such compositions in the treatment of patients suffering inflammatory, autoimmune and neurodegenerative diseases, for the prevention of ischemic injury, and for the preservation of organs that are to undergo a transplantation procedure.

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application is a continuation-in-part of U.S. applicationNo. 09/745,204 filed Dec. 19, 2000 and U.S. application No. 09/177,549filed Oct. 22, 1998, and claims the benefit of PCT Application No.PCT/US99/15074 filed Jul. 1, 1999 and Provisional Application No.60/091,689 filed Jul. 2, 1998 (each of which applications are herebyincorporated by reference in their entirety).

TECHNICAL FIELD

[0002] The present invention relates to novel classes of compounds whichare inhibitors of interleukin-1β converting enzyme and related proteases(“ICE/ced-3 family of cysteine proteases”), as well as pharmaceuticalcompositions comprising these compounds and to methods of using suchpharmaceutical compositions.

BACKGROUND OF THE INVENTION

[0003] Interleukin 1 (“IL-1”) is a major pro-inflammatory andimmunoregulatory protein that stimulates fibroblast differentiation andproliferation, the production of prostaglandins, collagenase andphospholipase by synovial cells and chondrocytes, basophil andeosinophil degranulation and neutrophil activation. Oppenheim, J. H. etal., Immunology Today, 7:45-56 (1986). As such, it is involved in thepathogenesis of chronic and acute inflammatory and autoimmune diseases.IL-1 is predominantly produced by peripheral blood monocytes as part ofthe inflammatory response. Mosely, B. S. et al., Proc. Nat. Acad. Sci.,84:4572-4576 (1987); Lonnemann, G. et al., Eur. J. Immunol.,19:1531-1536 (1989).

[0004] IL-1β is synthesized as a biologically inactive precursor,proIL-1p. ProIL-1β is cleaved by a cysteine protease calledinterleukin-1β converting enzyme (“ICE”) between Asp-116 and Ala-117 toproduce the biologically active C-terminal fragment found in human serumand synovial fluid. Sleath, P. R. et al., J. Biol. Chem.,265:14526-14528 (1992); A. D. Howard et al., J. Immunol., 147:2964-2969(1991).

[0005] ICE is a cysteine protease localized primarily in monocytes. Inaddition to promoting the pro-inflammatory and immunoregulatoryproperties of IL-1β, ICE, and particularly its homologues, also appearto be involved in the regulation of cell death or apoptosis. Yuan, J. etal., Cell, 75:641-652 (1993); Miura, M. et al., Cell, 75:653-660 (1993);Nett-Giordalisi, M. A. et al., J. Cell Biochem., 17B:117 (1993). Inparticular, ICE or ICE/ced-3 homologues are thought to be associatedwith the regulation of apoptosis in neurogenerative diseases, such asAlzheimer's and Parkinson's disease. Marx, J. and M. Baringa, Science,259:760-762 (1993); Gagliardini, V et al., Science, 263:826-828 (1994).

[0006] Thus, disease states in which inhibitors of the ICE/ced-3 familyof cysteine proteases may be useful as therapeutic agents include:infectious diseases, such as meningitis and salpingitis; septic shock,respiratory diseases; inflammatory conditions, such as arthritis,cholangitis, colitis, encephalitis, endocerolitis, hepatitis,pancreatitis and reperfusion injury, ischemic diseases such as themyocardial infarction, stroke and ischemic kidney disease; immune-baseddiseases, such as hypersensitivity; auto-immune diseases, such asmultiple sclerosis; bone diseases; and certain neurodegenerativediseases, such as Alzheimer's and Parkinson's disease. Such inhibitorsare also useful for the repopulation of hematopoietic cells followingchemo- and radiation therapy and for prolonging organ viability for usein transplantation.

[0007] ICE/ced-3 inhibitors represent a class of compounds useflil forthe control of the above-listed disease states. Peptide and peptidylinhibitors of ICE have been described. However, such inhibitors havebeen typically characterized by undesirable pharmacologic properties,such as poor oral absorption, poor stability and rapid metabolism.Plattner, J. J. and D. W. Norbeck, in Drug Discovery Technologies, C. R.Clark and W. H. Moos, Eds. (Ellis Horwood, Chichester, England, 1990),pp. 92-126. These undesirable properties have hampered their developmentinto effective drugs.

[0008] Accordingly, the need exists for compounds that can effectivelyinhibit the action of the ICE/ced-3 family of proteases, for use asagents for preventing unwanted apoptosis, and for treating chronic andacute forms of IL-1 mediated diseases such as inflammatory, autoimmuneor neurodegenerative diseases. The present invention satisfies this needand provides further related advantages.

SUMMARY OF THE INVENTION

[0009] In general, the compounds of this invention incorporate a(N-substituted)oxamyl group as a dipeptide mimetic. The resultingcompounds exhibit improved properties relative to their peptidiccounterparts, for example, such as improved cell penetration or improvedabsorption and metabolic stability resulting in enhancedbioavailability. This application claims priority from U.S. ProvisionalApplication No. 60/091,689, filed Jul. 2, 1998, and U.S. application No.09/177,549, filed Oct. 22, 1998 (both of which are hereby incorporatedby reference in their entirety).

[0010] One aspect of the instant invention is the compounds of theFormula I:

[0011] wherein A, B, R¹, R¹′ and R² are as defined below, as well aspharmaceutically acceptable salts thereof.

[0012] A further aspect of the instant invention is a pharmaceuticalcomposition comprising a compound of the above Formula I and apharmaceutically-acceptable carrier therefor.

[0013] Another aspect of this invention involves a method for treatingan autoimmune disease comprising administering an effective amount of apharmaceutical composition discussed above to a patient in need of suchtreatment.

[0014] Yet another aspect of the instant invention is a method fortreating an inflammatory disease comprising administering an effectiveamount of a pharmaceutical composition discussed above to a patient inneed of such treatment.

[0015] A further aspect of the instant invention is a method fortreating a neurodegenerative disease comprising administering aneffective amount of a pharmaceutical composition discussed above to apatient in need of such treatment.

[0016] Another aspect of the instant invention is a method of preventingischemic injury to a patient suffering from a disease associated withischemic injury comprising administering an effective amount of thepharmaceutical composition discussed above to a patient in need of suchtreatment.

[0017] A further aspect of the instant invention is a method forexpanding of hematopoietic cell populations and/or enhancing theirsurvival by contacting the cells with an effective amount of thepharmaceutical composition discussed above. Cell populations included inthe method of the invention include (but are not limited to)granulocytes, monocytes, erthrocytes, lymphocytes and platelets for usein cell transfusions.

[0018] An alternate aspect of the instant invention is a method ofprolonging the viability of an organ that has been removed from thedonor for the purpose of a future transplantation procedure, whichcomprises applying an effective amount of the pharmaceutical compositiondiscussed above to the organ, thereby prolonging the viability of theorgan as compared to an untreated organ. The organ may be an intactorgan, or isolated cells derived from an organ (e.g., isolatedpancreatic islet cells, isolated dopaminergic neurons, blood orhematopoietic cells).

[0019] These and other aspects of this invention will be evident uponreference to the following detailed description.

DETAILED DESCRIPTION OF THE INVENTION

[0020] As mentioned above, one aspect of the instant invention is thecompounds of the Formula I:

[0021] wherein:

[0022] A is a natural or unnatural amino acid of Formula IIa-i:

[0023] B is a hydrogen atom, a deuterium atom, alkyl, cycloalkyl,phenyl, substituted phenyl, naphthyl, substituted naphtbyl,2-benzoxazolyl, substituted 2-oxazolyl, (CH₂)_(n)cycloalkyl,(CH₂)_(n)phenyl, (CH₂)_(n)(substituted phenyl), (CH₂)_(n)(1 or2-naphthyl), (CH₂)_(n)(substituted 1 or 2-naphthyl),(CH₂)_(n)(heteroaryl), (CH₂)_(n)(substituted heteroaryl), halomethyl,CO₂R¹², CONR¹³R¹⁴, CH₂ZR¹⁵, CH₂OCO(aryl), CH₂OCO(heteroaryl), orCH₂OPO(R¹⁶)R¹⁷, where Z is an oxygen or a sulfur atom, or B is a groupof the Formula IlIa-c:

[0024] R¹ is alkyl, cycloalkyl, substituted cycloalkyl,(cycloalkyl)alkyl, substituted (cycloalkyl)alkyl, phenyl, substitutedphenyl, phenylalkyl, substituted phenylalkyl, naphthyl, substitutednaphthyl, (1 or 2 naphthyl)alkyl, substituted (1 or 2 naphthyl)alkyl,heterocycle, substituted heterocycle, (heterocycle)alkyl, substituted(heterocycle)alkyl, R^(1a)(R^(1b))N, or R^(1c)O;

[0025] R¹′ is hydrogen, alkyl, phenyl, substituted phenyl, naphthyl,substituted naphthyl, heterocycle or substituted heterocycle;

[0026] or R¹ and R¹′ taken together with the nitrogen atom to which theyare attached form a heterocycle or substituted heterocycle;

[0027] R² is hydrogen, lower alkyl, cycloalkyl, (cycloalkyl)aLkyl,phenyl, substituted phenyl, phenylalkyl, substituted phenylalkyl,naphthyl, substituted naphthyl, (1 or 2 naphthyl)alkyl, or substituted(1 or 2 naphthyl)alkyl;

[0028] and wherein:

[0029] R^(1a) and R^(1b) are independently hydrogen, alkyl, cycloalkyl,(cycloalkyl)alkyl, phenyl, substituted phenyl, phenylalkyl, substitutedphenylalkyl, naphthyl, substituted naphthyl, (1 or 2 naphthyl)alkyl,substituted (1 or 2 naphthyl)alkyl, beteroaryl, substituted heteroaryl,(heteroaryl)alkyl, or substituted (heteroaryl)alkyl, with the provisothat R^(1a) and R^(1b) cannot both be hydrogen;

[0030] R^(1c) is alkyl, cycloalkyl, (cycloalkyl)alkyl, phenyl,substituted phenyl, phenylalkyl, substituted phenylalkyl, naphthyl,substituted naphthyl, (1 or 2 naphthyl)alkyl, substituted (1 or 2naphthyl)alkyl, heteroaryl, substituted heteroaryl, (heteroaryl)alkyl,or substituted (heteroaryl)alkyl;

[0031] R³ is C₁₋₆ lower alkyl, cycloalkyl, phenyl, substituted phenyl,(CH₂)_(n)NH₂, (CH₂)_(n)NHCOR⁹, (CH₂)_(n)N(C═NH)NH₂, (CH₂)_(m)CO₂R²,(CH₂)_(m)OR¹⁰, (CH₂)_(m)SR¹¹, (CH₂)_(n)cycloalkyl, (CH₂)_(n)phenyl,(CH₂)_(n)(substituted phenyl), (CH₂)_(n)(1 or 2-naphthyl) or(CH₂)_(n)(heteroaryl), wherein heteroaryl includes pyridyl, thienyl,furyl, thiazolyl, imidazolyl, pyrazolyl, isoxazolyl, pyrazinyl,pyrimidyl, triazinyl, tetrazolyl, and indolyl;

[0032] R^(3a) is hydrogen or methyl, or R³ and R^(3a) taken together are—(CH₂)_(d)— where d is an interger from 2 to 6;

[0033] R⁴ is phenyl, substituted phenyl, (CH₂)_(m)phenyl,(CH₂)_(m)(substituted phenyl), cycloalkyl, or benzofused cycloalkyl;

[0034] R⁵ is hydrogen, lower alkyl, cycloalkyl, phenyl, substitutedphenyl, (CH₂)_(n)cycloalkyl, (CH₂)_(n)phenyl, (CH₂)_(n)(substitutedphenyl), or (CH₂)_(n)(l or 2-naphthyl);

[0035] R⁶ is hydrogen, fluorine, oxo, lower alkyl, cycloalkyl, phenyl,substituted phenyl, naphthyl, (CH₂)_(n)cycloalkyl, (CH₂)_(n)phenyl,(CH₂)_(n)(substituted phenyl), (CH₂)_(n)(1 or 2-naphthyl), OR¹⁰, SR¹¹ orNHCOR⁹;

[0036] R⁷ is hydrogen, oxo (i.e., ═O), lower alkyl, cycloalkyl, phenyl,substituted phenyl, naphthyl, (CH₂)_(n)cycloalkyl, (CH₂)_(n)phenyl,(CH₂)_(n)(substituted phenyl), or (CH₂)_(n)(1 or 2-naphthyl);

[0037] R⁸ is lower alkyl, cycloalkyl, (CH₂)_(n)cycloalkyl,(CH₂)_(n)phenyl, (CH₂)_(n)(substituted phenyl), (CH₂)_(n)(1 or2-naphthyl), or COR⁹;

[0038] R⁹ is hydrogen, lower alkyl, cycloalkyl, phenyl, substitutedphenyl, naphthyl, (CH₂)_(n)cycloalkyl, (CH₂)_(n)phenyl,(CH₂)_(n)(substituted phenyl), (CH₂)_(n)(1 or 2-naphthyl), OR¹², orNR¹³R¹⁴;

[0039] R¹⁰ is hydrogen, lower alkyl, cycloalkyl, phenyl, substitutedphenyl, naphthyl, (CH₂)_(n)cycloalkyl, (CH₂)_(n)phenyl,(CH₂)_(n)(substituted phenyl), or (CH₂)_(n)(1 or 2-naphthyl);

[0040] R¹¹ is lower alkyl, cycloalkyl, phenyl, substituted phenyl,naphthyl, (CH₂)_(n)cycloalkyl, (CH₂)_(n)phenyl, (CH₂)_(n)(substitutedphenyl), or (CH₂)_(n)(1 or 2-naphthyl);

[0041] R¹² is lower alkyl, cycloalkyl, (CH₂)_(n)cycloalkyl,(CH₂)_(n)phenyl, (CH₂)_(n)(substituted phenyl), or (CH₂)_(n)(1 or2-naphthyl);

[0042] R¹³ is hydrogen, lower alkyl, cycloalkyl, phenyl, substitutedphenyl, naphthyl, substituted naphthyl, (CH₂)_(n)cycloalkyl,(CH₂)_(n)phenyl, (CH₂)_(n)(substituted phenyl), or (CH₂)_(n)(1 or2-naphthyl);

[0043] R¹⁴ is hydrogen or lower alkyl;

[0044] or R¹³ and R¹⁴ taken together form a five to seven memberedcarbocyclic or heterocyclic ring, such as morpholine, or N-substitutedpiperazine;

[0045] R¹⁵ is phenyl, substituted phenyl, napbthyl, substitutednaphthyl, heteroaryl, (CH₂)_(n)phenyl, (CH₂)_(n)(substituted phenyl),(CH₂)_(n)(1 or 2-naphthyl), or (CH₂)_(n)(heteroaryl);

[0046] R¹⁶ and R¹⁷ are independently lower alkyl, cycloalkyl, phenyl,substituted phenyl, naphthyl, phenylalkyl, substituted phenylalkyl, or(cycloalkyl)alkyl;

[0047] R¹⁸ and R¹⁹ are independently hydrogen, alkyl, phenyl,substituted phenyl, (CH₂)_(n)phenyl, (CH₂)_(n)(substituted phenyl), orR¹⁸ and R¹⁹ taken together are —(CH═CH)₂—;

[0048] R²⁰ is hydrogen, alkyl, phenyl, substituted phenyl,(CH₂)_(n)phenyl, (CH₂)_(n)(substituted phenyl);

[0049] R²¹ , R²² and R²³ are independently hydrogen, or alkyl;

[0050] X is CH₂, (CH₂)₂, (CH₂)₃, or S;

[0051] Y¹ is O or NR²³;

[0052] Y² is CH₂, O, or NR²³;

[0053] a is 0 or 1;

[0054] b is 1 or 2, provided that when a is 1 then b is 1

[0055] c is 1 or 2, provided that when c is 1 then a is 0 and b is 1;

[0056] m is 1 or 2; and

[0057] n is 1, 2, 3 or 4;

[0058] or a pharmaceutically acceptable salt thereof

[0059] As used herein, the term “alkyl” means a straight or branched C₁to C₁₀ carbon chain, such as methyl, ethyl, tert-butyl, iso-propyl,n-octyl, and the like. The term “lower alkyl” means a straight chain orbranched C₁ to C₆ carbon chain, such as methyl, ethyl, iso-propyl, andthe like.

[0060] The term “cycloalkyl” means a mono-, bi-, or tricyclic ring thatis either fully saturated, partially of fully unsaturated, or aromatic.Examples of such a ring include cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, cycloheptyl, adamantyl, cyclooctyl, cis- or trans decalin,bicyclo[2.2.1]hept-2-ene, cyclohex-1-enyl, cyclopent-1-enyl,1,4-cyclooctadienyl, and the like.

[0061] The term “(cycloalkyl)alkyl” means the above-defined alkyl groupsubstituted with one of the above cycloalkyl rings. Examples of such agroup include (cyclohexyl)methyl, 3-(cyclopropyl)-n-propyl,5-(cyclopentyl)hexyl, 6-(adamantyl)hexyl, and the like.

[0062] The term “substituted phenyl” specifies a phenyl groupsubstituted with one or more substituents chosen from halogen, hydroxy,protected hydroxy, cyano, nitro, trifluoromethyl, alkyl, alkoxy, acyl,acyloxy, carboxy, protected carboxy, carboxymethyl, protectedcarboxymethyl, hydroxymethyl, protected hydroxymethyl, amino, protectedamino, (monosubstituted)amino, protected (monosubstituted)amino,(disubstituted)amino, carboxamide, protected carboxamide, N-(loweralkyl)carboxamide, protected N-(lower alkyl)carboxamide, N,N-di(loweralkyl)carboxamide, N-((lower alkyl)sulfonyl)amino,N-(phenylsulfonyl)amino, substituted or unsubstituted heterocycle,substituted or unsubstituted cycloalkyl, or by a substituted orunsubstituted phenyl group, such that in the latter case a biphenyl ornaphthyl group results, or wherein two adjacent allkyl substituents onthe substituted phenyl ring taken together form a cycloalkyl to yield,for example, tetrahydronaphthyl or indanyl.

[0063] Examples of the term “substituted phenyl” includes a mono-, di-,tri-, tetra- or penta(halo)phenyl group such as 2-, 3- or4-chlorophenyl, 2,6-dichlorophenyl, 2,5-dichlorophenyl,3,4-dichlorophenyl, 2-,3- or 4-bromophenyl, 3,4-dibromophenyl,3-chloro-4-fluorophenyl, 2-, 3- or 4-fluorophenyl,2,4,6-trifluorophenyl, 2,3,5,6-tetrafluorophenyl,2,3,4,5-tetrafluorophenyl, 2,3,4,5,6-pentafluorophenyl, and the like; amono or di(hydroxy)phenyl group such as 2-, 3-, or 4-hydroxyphenyl,2,4-dihydroxyphenyl, the protected-hydroxy derivatives thereof and thelike; a nitrophenyl group such as 2-, 3-, or 4-nitrophenyl; acyanophenyl group, for example, 2-,3- or 4-cyanophenyl; a mono- ordi(alkyl)phenyl group such as 2-, 3-, or 4-methylphenyl,2,4-dimethylphenyl, 2-, 3- or 4-(iso-propyl)phenyl, 2-, 3-, or4-ethylphenyl, 2-, 3- or 4-(n-propyl)phenyl and the like; a mono ordi(alkoxy)phenyl group, for example, 2,6-dimethoxyphenyl, 2-, 3- or4-(iso-propoxy)phenyl, 2-, 3- or 4-(t-butoxy)phenyl,3-ethoxy-4-methoxyphenyl and the like; 2-, 3- or4-trifluoromethylphenyl; a mono- or dicarboxyphenyl or (protectedcarboxy)phenyl group such as 2-, 3- or 4-carboxyphenyl or2,4-di(protected carboxy)phenyl; a mono- or di(hydroxymethyl)phenyl or(protected hydroxymethyl)phenyl such as 2-, 3- or 4-(protectedhydroxymethyl)phenyl or 3,4-di(hydroxymethyl)phenyl; a mono- ordi(aminomethyl)phenyl or (protected aminomethyl)phenyl such as 2-, 3- or4-(aminomethyl)phenyl or 2,4-(protected aminomethyl)phenyl; or a mono-or di(N-(methylsulfonylamino))phenyl such as 2, 3 or4-(N-(methylsulfonylamino))phenyl. Also, the term “substituted phenyl”represents disubstituted phenyl groups wherein the substituents aredifferent, for example, 3-methyl-4-hydroxyphenyl,3-chloro-4-hydroxyphenyl, 2-methoy-4-bromophenyl,4-ethyl-2-hydroxyphenyl, 3-hydroxy-4-nitrophenyl,2-hydroxy-4-chlorophenyl, and the like.

[0064] The term “phenylalkyl” means one of the above phenyl groupsattached to one of the above-described alkyl groups, and the term“substituted phenylalkyl” means that either the phenyl or the alkyl, orboth, are substituted with one or more of the above-definedsubstituents. Examples of such groups include 2-phenyl-1-chloroethyl,2-(4′-methoxyphenyl)ethyl, 4-(2′,6′-dihydroxy phenyl)n-hexyl,2-(5′-cyano-3′-methoxyphenyl)n-pentyl, 3-(2′,6′-dimethylphenyl)n-propyl,4-chloro-3-aminobenzyl, 6-(4′-methoxyphenyl)-3-carboxy(n-hexyl),5-(4′-aminomethylphenyl)-3-(aminomethyl)n-pentyl,5-phenyl-3-oxo-n-pent-1-yl, (4-hydroxynapth-2-yl)methyl, and the like.

[0065] The term “substituted naphthyl” means a naphthyl groupsubstituted with one or more of the above-identified substituents, andthe term “(1 or 2 naphthyl)alkyl” means a naphthyl (1 or 2) attached toone of the above-described alkyl groups.

[0066] The terms “halo” and “halogen” refer to the fluoro, chloiro,bromo or iodo groups. These terms may also be used to describe one ormore halogens, which are the same or different. Preferred halogens inthe context of this invention are chloro and fluoro.

[0067] The term “aryl” refers to aromatic five and six memberedcarbocyclic rings. Six membered rings are preferred.

[0068] The term “heterocycle” denotes optionally substitutedfive-membered or six-membered heterocyclic rings that have 1 to 4heteroatoms, such as oxygen, sulfur and/or nitrogen atoms, in particularnitrogen, either alone or in conjunction with sulfur or oxygen ringatoms, and include aromatic heterocycles (also referred to herein as“heteroaryls”). The following ring systems are representative examplesof aromatic heterocyclic radicals denoted by the term heteroaryl(whether substituted or unsubstituted): thienyl, furyl, pyrrolyl,pyrrolidinyl, imidazolyl, isoxazolyl, triazolyl, thiadiazolyl,oxadiazolyl, tetrazolyl, thiatriazolyl, oxatriazolyl, pyridyl,pyrimidyl, pyrazinyl, pyridazinyl, oxazinyl, triazinyl, thiadiazinyltetrazolo, 1,5-[b]pyridazinyl and purinyl, as well as benzo-fusedderivatives, for example, benzoxazolyl, benzothiazolyl, benzimidazolyland indolyl. Non-aromatic heterocycles include, for exatriple,morpholinyl, pyrrolidinonyl, pyrrolidinyl, piperidinyl, piperazinyl,hydantoinyl, valerolactamyl, oxiranyl, oxetanyl, tetrahydrofuranyl,tetrahydropyranyl, tetrahydropyridinyl, tetrahydroprimidinyl,tetrahydrothiophenyl, tetrahydrothiopyranyl, tetrahydropyrimidinyl,tetrahydrothiophenyl, tetrahydrothiopyranyl, and the like.

[0069] Substituents for the above optionally substituted cycloalkyl orheterocycle rings are as listed above for substituted phenyl, and morespecifically include from one to three halo, trihalomethyl, amino,protected amino, amino salts, mono-substituted amino, di-substitutedamino, carboxy, protected carboxy, carboxylate salts, hydroxy, protectedhydroxy, salts of a hydroxy group, lower alkoxy, lower alkylthio, loweralkyl, substituted lower alkyl, cycloalkyl, substituted cycloalkyl,(cycloalkyl)alkyl, substituted (cycloalkyl)alkyl, phenyl, substitutedphenyl, phenylalkyl, and substituted phenylalkyl groups. More than onesubstituent may be made at any given atom of the heterocyclic ring,including carbocyclic or heterocyclic substituents that form a spirounion. “Trihalomethyl” can be trifluoromethyl, trichloromethyl,tribromomethyl or triiodomethyl, “lower alkoxy” means a C₁ to C₄ alkoxygroup, similarly, “lower alkylthio” means a C₁ to C₄ alkylthio group.The term “substituted lower alkyl” means the above-defined lower alkylgroup substituted from one to three times by a hydroxy, protectedhydroxy, amino, protected amino, cyano, halo, trifluoromethyl,mono-substituted amino, di-substituted amino, lower alkoxy, loweralkylthio, carboxy, protected carboxy, or a carboxy, amino, and/orhydroxy salt.

[0070] The terms “substituted (cycloalkyl)alkyl” and “substitutedcycloalkyl” are as defined above with regard to (cycloalkyl)alkyl andcycloalkyl, but substituted with one or more substituents as identifiedabove with regard to substituted phenyl, cycloalkyl and/or heterocycle.The term “(monosubstituted)amino” refers to an amino group with onesubstituent chosen from the group consisting of phenyl, substitutedphenyl, alkyl, substituted alkyl, C₁ to C₇ acyl, C₂ to C₇ alkenyl, C₂ toC₇ substituted alkenyl, C₂ to C₇ alkynyl, C₇ to C₁₆ alkylaryl, C₇ to C₁₆substituted alkylaryl and heteroaryl group. The (monosubstituted)aminocan additionally have an amino-protecting group as encompassed by theterm “protected (monosubstituted)amino.” The term “(disubstituted)amino”refers to amino groups with two substituents chosen from the groupconsisting of phenyl, substituted phenyl, alkyl, substituted alkyl, C₁to C₇ acyl, C₂ to C₇ alkenyl, C₂ to C₇ alkynyl, C₇ to C₁₆ alkylaryl, C₇to C₁₆ substituted alkylaryl and heteroaryl. The two substituents can bethe same or different. The term “heteroaryl(alkyl)” denotes an alkylgroup as defmed above, substituted at any position by a heteroarylgroup, as above defined.

[0071] More specifically, and in addition to the substituents disclosedabove, the term “substituted” as used herein means a chemical moietywherein at least one hydrogen atom is replaced with a substituent. Inthe case of a keto substituent (“C(═O)”) two hydrogen atoms arereplaced. Substituents include halogen, hydroxy, alkyl, substitutedalkyl (such as haloalkyl, mono- or di-substituted aminoalkyl,alkyloxyalkyl, and the like), aryl, substituted aryl, arylalkyl,substituted arylalkyl, heterocycle, substituted heterocycle,heterocyclealkyl, substituted heterocyclealkyl, —NR_(a)R_(b),NR_(a)C(═O)R_(b), —NR_(a)C(═O)NR_(a)R_(b), —NR_(a)C(═O)OR_(b)NR_(a)SO₂R_(b), —OR_(a), —C(═O)R_(a) —C(═O)OR_(a) C(═O)NR_(a)R_(b),—OC(═O)R_(a), —OC(═O)OR_(a), —OC(═O)NR_(a)R_(b), —NR_(a)SO₂R_(b), or aradical of the formula —Y—Z—R_(a) where Y is alkanediyl, substitutealkanediyl, or a direct bond, Z is —O—, —S—, —S(═O)—, —S(═O)₂—,—N(R_(b))—, —C(═O)—, —C(═O)O—, —OC(═O)—, —N(R_(b))C(═O)—,—C(═O)N(R_(b))— or a direct bond, wherein R_(a) and R_(b) are the sameor different and independently hydrogen, amino, alkyl, substituted alkyl(including halogenated alkyl), aryl, substituted aryl, arylalkyl,substituted arylalkyl, heterocycle, substituted heterocycle,heterocylealkyl or substituted heterocyclealkyl, or wherein R_(a) andR_(b) taken together with the nitrogen atom to which they are attachedform a heterocycle or substituted heterocycle.

[0072] Furthermore, the above optionally substituted five-membered orsix-membered heterocyclic rings, and the above cycloalkyl rings, canoptionally be fused to an aromatic 5-membered or 6-membered aryl,carbocyclic or heterocyclic ring system. For example, the rings can beoptionally fused to an aromatic 5-membered or 6-membered ring systemsuch as a pyridine or a triazole system, and preferably to a benzenering.

[0073] The term “pharmaceutically-acceptable salt” encompasses thosesalts that form with the carboxylate anions and includes salts formedwith the organic and inorganic cations such as those chosen from thealkali and alkaline earth metals, (for example, lithium, sodium,potassium, magnesium, barium and calcium); and ammonium ion; and theorganic cations (for example, dibenzylammonium, benzylammonium,2-hydroxyethylammonium, bis(2-hydroxyethyl)ammonium,phenylethylbenzyl-ammonium, dibenzylethylenediammonium, and likecations.) Other cations encompassed by the above term include theprotonated form of procaine, quinine and N-methylglucosamine, theprotonated forms of basic amino acids such as glycine, ornithine,histidine, phenylglycine, lysine, and arginine. Furthermore, anyzwitterionic form of the instant compounds formed by a carboxylic acidand an amino group is referred to by this term. A preferred cation forthe carboxylate anion is the sodium cation. Furthermore, the termincludes salts that form by standard acid-base reactions with basicgroups (such as amino groups) and includes organic or inorganic acids.Such acids include hydrochloric, sulfuric, phosphoric, acetic, succinic,citric, lactic, maleic, fumaric, palmitic, cholic, pamoic, mucic,D-glutamic, D-camphoric, glutaric, phthalic, tartaric, lauric, stearic,salicyclic, methanesulfonic, benzenesulfonic, sorbic, picric, benzoic,cinnamic, and the like acids.

[0074] The compounds of Formula I may also exist as solvates andhydrates. Thus, these compounds may crystallize with, for example,waters of hydration, or one, a number of, or any fraction thereof ofmolecules of the mother liquor solvent. The solvates and hydrates ofsuch compounds are included within the scope of this invention.

[0075] The term “carboxy-protecting group” as used herein refers to oneof the ester derivatives of the carboxylic acid group commonly employedto block or protect the carboxylic acid group while reactions arecarried out on other functional groups on the compound. Examples of suchcarboxylic acid protecting groups include t-butyl, 4-nitrobenzyl,4-methoxybenzyl, 3,4-dimethoxybenzyl, 2,4-dimethoxybenzyl,2,4,6-trimethoxybenzyl, 2,4,6-trimethylbenzyl, pentamethylbenzyl,3,4-methylenedioxybenzyl, benzhydryl, 4,4′-dimethoxytrityl,4,4′,4″-trimethoxytrityl, 2-phenylpropyt, trimethylsilyl,t-butyldimethylsilyl, phenacyl, 2,2,2-trichloroethyl,O-(trimethylsilyl)ethyl, P-(di(n-butyl)methylsilyl)ethyl,p-toluenesulfonylethyl, 4-nitrobenzylsulfonylethyl, allyl, cinnamyl,1-(trimethylsilylmethyl)-propenyl and like moieties. The species ofcarboxy-protecting group employed is not critical so long as thederivatized carboxylic acid is stable to the conditions of subsequentreaction(s) and can be removed at the appropriate point withoutdisrupting the remainder of the molecule. Further examples of thesegroups are found in C. B. Reese and E. Haslam, “Protective Groups inOrganic Chemistry,”0 J. G. W McOmie, Ed., Plenum Press, New York, N.Y.,1973, Chapter 5, respectively, and T. W Greene and P. G. M. Wuts,“Protective Groups in Organic Synthesis,” 2nd ed., John Wiley and Sons,New York, N.Y., 1991, Chapter 5, each of which is incorporated herein byreference. A related term is “protected carboxy,” which refers to acarboxy group substituted with one of the above carboxy-protectinggroups.

[0076] The term “hydroxy-protecting group” refers to readily cleavablegroups bonded to hydroxyl groups, such as the tetrahydropyranyl,2-metboxyprop-2-yl, 1-ethoxyeth-1-yl, methoxymethyl,β-methoxyethoxymethyl, methylthiomethyl, t-butyl, t-amyl, trityl,4-methoxytrityl, 4,4′-dimethoxytrityl, 4,4′,4″-trimethoxytrityl, benzyl,allyl, trimethylsilyl, (t-butyl)dimethylsilyl,2,2,2-trichloroethoxycarbonyl, and the like.

[0077] Further examples of hydroxy-protecting groups are described by C.B. Reese and E. Haslam, “Protective Groups in Organic Chemistry,” J. G.W McOmie, Ed., Plenum Press, New York, N.Y., 1973, Chapters 3 and 4,respectively, and T. W Greene and P. G. M. Wuts, “Protective Groups inOrganic Synthesis,” Second Edition, John Wiley and Sons, New York, N.Y.,1991, Chapters 2 and 3. A preferred hydroxy-protecting group is thetert-butyl group. The related term “protected hydroxy” denotes a hydroxygroup bonded to one of the above hydroxy-protecting groups.

[0078] The term “amino-protecting group” as used herein refers tosubstituents of the amino group commonly employed to block or protectthe amino functionality while reacting other functional groups of themolecule. The term “protected (monosubstituted)amino” means there is anamino-protecting group on the monosubstituted amino nitrogen atom.

[0079] Examples of such amino-protecting groups include the formyl(“For”) group, the trityl group, the phthalimido group, thetrichloroacetyl group, the trifluoroacetyl group, the chloroacetyl,bromoacetyl, and iodoacetyl groups, urethane-type protecting groups,such as t-butoxycarbonyl (“Boc”), 2-(4-biphenylyl)propyl-2-oxycarbonyl(“Bpoc”), 2-phenylpropyl-2-oxycarbonyl (“Poc”),2-(4-xenyl)isopropoxycarbonyl, 1, 1-diphenylethyl-1-oxycarbonyl,1,-diphenylpropyl-1-oxycarbonyl,2-(3,5-dimethoxyphenyl)propyl-2-oxycarbonyl (“Ddz”),2-(p-toluyl)propyl-2-oxycarbonyl, cyclopentanyloxycarbonyl, I-methylcyclopentanyl-oxycarbonyl, cyclohexanyloxy-carbonyl,1-methyl-cyclohexanyloxy-carbonyl, 2-methylcyclohexanyl-oxycarbonyl,2-(4-toluylsulfonyl)ethoxycarbonyl, 2-(methylsulfonyl)ethoxycarbonyl,2-(triphenylphosphino)-ethoxycarbonyl, 9-fluorenyl-methoxycarbonyl(“Fmoc”), 2-(trimethylsilyl)ethoxycarbonyl, allyloxycarbonyl,1-(trimethylsilylmethyl)prop-1-enyloxycarbonyl,5-benzisoxalylmethoxycarbonyl, 4-acetoxybenzyl-oxycarbonyl,2,2,2-trichloroethoxycarbonyl, 2-ethynyl-2-propoxycarbonyl,cyclopropylmethoxycarbonyl, isobornyloxycarbonyl,1-piperidyloxycarbonyl, benzyl-oxycarbonyl (“Cbz”),4-phenylbenzyloxycarbonyl, 2-methylbenzyloxycarbonyl,α-2,4,5,-tetramethylbenzyl-oxycarbonyl (“Tmz”),4-methoxybenzyloxycarbonyl, 4-fluorobenzyl-oxycarbonyl,4-chlorobenzyloxycarbonyl, 3-chlorobenzyloxycarbonyl,2-chlorobenzyloxy-carbonyl, 2,4-dichlorobenzyloxycarbonyl,4-bromobenzyloxycarbonyl, 3-bromobenz yloxy-carbonyl,4-nitrobenzyloxycarbonyl, 4-cyanobenzyloxycarbonyl,4-(decyloxy)benzyloxy-carbonyl and the like; the benzoylmethylsulfonylgroup, the 2,2,5,7,8-pentamethylchroman-6-sulfonyl group (“PMC”), thedithiasuccinoyl (“Dts”) group, the 2-(nitro)phenyl-sulfenyl group(“Nps”), the diphenylphosphine oxide group, and like amino-protectinggroups. The species of amino-protecting group employed is not criticalso long as the derivatized amino group is stable to the conditions ofthe subsequent reaction(s) and can be removed at the appropriate pointwithout disrupting the remainder of the molecule. Preferred amino-protecting groups are Boc, Cbz and Fmoc. Further examples ofamino-protecting groups embraced by the above term are well known inorganic synthesis and the peptide art and are described by, for example,T. W. Greene and P. G. M. Wuts, “Protective Groups in OrganicSynthesis,” 2nd ed., John Wiley and Sons, New York, N.Y., 1991, Chapter7, M. Bodanzsky, “Principles of Peptide Synthesis,” 1st and 2nd revisedEd., Springer-Verlag, New York, N.Y., 1984 and 1993, and J. M. Stewartand J. D. Young, “Solid Phase Peptide Synthesis,” 2nd Ed., PierceChemical Co., Rockford, Ill., 1984, E. Atherton and R. C. Shephard,“Solid Phase Peptide Synthesis—A Practical Approach” IRL Press, Oxford,England (1989), each of which is incorporated herein by reference. Therelated term “protected amino” defines an amino group substituted withan amino-protecting group discussed above.

[0080] The terms “natural and unnatural amino acid” refers to both thenaturally occurring amino acids and other non-proteinogenic α-aminoacids commonly utilized by those in the peptide chemistry arts whenpreparing synthetic analogues of naturally occurring peptides, includingD and L forms. The naturally occurring amino acids are glycine, alanine,valine, leucine, isoleucine, serine, methionine, threonine,phenylalanine, tyrosine, tryptophan, cysteine, proline, histidine,aspartic acid, asparagine, glutamic acid, glutamine, y-carboxyglutamicacid, arginine, ornithine and lysine. Examples of unnatural alpha-aminoacids include hydroxylysine, citrulline, kynurenine,(4-aminophenyl)alanine, 3-(2′-naphthyl)alanine, 3-(1′-naphthyl)alanine,methionine sulfone, (t-butyl)alanine, (t-butyl)glycine,4-hydroxyphenyl-glycine, aminoalanine, phenylglycine, vinylalanine,propargyl-gylcine, 1,2,4-triazolo-3-alanine, thyronine,6-hydroxytryptophan, 5-hydroxytryptophan, 3-hydroxy-kynurenine,3-aminotyrosine, trifluoromethylalanine, 2-thienylalanine,(2-(4-pyridyl)ethyl)cysteine, 3,4-dimethoxy-phenylalanine,3-(2′-thiazolyl)alanine, ibotenic acid,1-amino-1-cyclopentane-carboxylic acid, 1-amino-1-cyclohexanecarboxylicacid, quisqualic acid, 3-(trifluoromethylphenyl)alanine,(cyclohexyl)glycine, thiohistidine, 3-methoxytyrosine, norleucine,norvaline, alloisoleucine, homoarginine, thioproline, dehydro-proline,hydroxyproline, homoproline, indoline-2-carboxylic acid,1,2,3,4-tetrahydroisoquinoline-3-carboxylic acid,1,2,3,4-tetrahydroquinoline-2-carboxylic acid, a-amino-n-butyric acid,cyclohexylalanine, 2-amino-3-phenylbutyric acid, phenylalaninesubstituted at the ortho, meta, or para position of the phenyl moietywith one or two of the following groups: a (C₁ to C₄)alkyl, a (C₁ toC₄)alkoxy, a halogen or a nitro group, or substituted once with amethylenedioxy group; β-2- and 3-thienylalanine; β-2- and3-furanylalanine; β-2-, 3- and 4-pyridylalanine; β-(benzothienyl-2- and3-yl)alanine; β-(1- and 2-naphthyl)alanine; O-alkylated derivatives ofserine, threonine or tyrosine; S-alkylated cysteine, S-alkylatedhomocysteine, the O-sulfate, O-phosphate and O-carboxylate esters oftyrosine; 3-(sulfo)tyrosine, 3-(carboxy)tyrosine, 3-(phospho)tyrosine,the 4-methane-sulfonic acid ester of tyrosine, 4-methanephosphonic acidester of tyrosine, 3,5-diiodotyrosine, 3-nitrotyrosine, ε-alkyllysine,and delta-alkyl ornithine. Any of these α-amino acids may be substitutedwith a methyl group at the alpha position, a halogen at any position ofthe aromatic residue on the a-amino side chain, or an appropriateprotective group at the O, N, or S atoms of the side chain residues.Appropriate protective groups are discussed above.

[0081] Depending on the choice of solvent and other conditions known tothe practitioner skilled in the art, compounds of this invention mayalso take the ketal or acetal form, which forms are included in theinstant invention. In particular, when R² is hydrogen compounds ofFormula Ia may exist in the cyclic ketal or acetal form Formula Ia′shown below:

[0082] Similarly, when R² of Formula I is a moiety other than hydrogen,and depending upon the choice of solvents as noted above (e.g., R²OH),the compounds of the cyclic ketal or acetal form include compoundshaving Formula Ia″ as shown below.

[0083] In addition, it should be understood that the equilibrium formsof the compounds of this invention may include tautomeric forms. Allsuch forms of these compounds are expressly included in the presentinvention.

[0084] The compounds of this invention may be modified by appropriatefunctionalities to enhance selective biological properties. Suchmodifications are known in the art and include those which increasebiological penetration into a given biological system (e.g., blood,lymphatic system, central nervous system), increase oral availability,increase solubility to allow administration by injection, altermetabolism and alter rate of exertion. In addition, the compounds may bealtered to pro-drug form such that the desired compound is created inthe body of the patient as the result of the action of metabolic orother biochemical processes on the pro-drug. Some examples of pro-drugforms include ketal, acetal, oxime, and hydrazone forms of compoundswhich contain ketone or aldehyde groups, especially where they occur inthe group donated as “A” in Formula I or the modified aspartic acidresidue attached to the group denoted as “A”.

[0085] Compounds of this invention with respect to the group “R¹” inFormula I, include those wherein:

[0086] R¹ is phenyl, substituted phenyl, phenylalkyl, substitutedphenylalkyl, naphthyl, substituted naphthyl, (1 or 2 naphthyl)alkyl,heteroaryl, or (heteroaryl)alkyl.

[0087] More typically, the compounds of this invention with respect tothe group “R¹” include those wherein:

[0088] R¹ is phenyl, substituted phenyl, phenylalkyl, substitutedphenylalkyl, naphthyl, substituted naphthyl, or (1 or 2 naphthyl)alkyl.

[0089] Compounds of this invention with respect to the group “R¹′”include those wherein:

[0090] R¹′ is hydrogen, lower alkyl and aryl.

[0091] Compounds of this invention with respect to the group “A” inFormula I, include those of Formula IIa wherein:

[0092] R³ is lower alkyl, cycloalkyl, phenyl, substituted phenyl,(CH₂)_(n)NH₂, (CH₂)_(m)OR¹⁰, (CH₂)_(m)SR¹¹,(CH₂)_(n)cycloalkyl,(CH₂)_(n)phenyl, (CH₂)_(n)(substituted phenyl), or(CH₂)_(n)(1 or 2-naphthyl);

[0093] R^(3a) is hydrogen;

[0094] R¹⁰ is hydrogen, lower alkyl, cycloalkyl, phenyl, substitutedphenyl, naphthyl, (CH₂)_(n)cycloalkyl, (CH₂)_(n)phenyl,(CH₂)_(n)(substituted phenyl), or (CH₂)_(n)(1 or 2-naphthyl);

[0095] R¹¹ is lower alkyl, cycloalkyl, phenyl, substituted phenyl,naphthyl, (CH₂)_(n)cycloalkyl, (CH₂)_(n)phenyl, (CH₂)_(n)(substitutedphenyl), or (CH₂)_(n)(1 or 2-naphthyl); and

[0096] n=1-4 and m=1 or 2.

[0097] Compounds of this invention with respect to the group “A” inFormula I, also include those of Formula IIb wherein:

[0098] R⁴ is phenyl, substituted phenyl, (CH₂)_(m)phenyl,(CH₂)_(m)(substituted phenyl), cycloalkyl, or 2-indanyl; and

[0099] m=1 or 2.

[0100] Another group of compounds with respect to the group “A” inFormula I, include those of Formula IId wherein:

[0101] R⁶ is hydrogen, fluorine, cycloalkyl, phenyl, substituted phenyl,naphthyl, (CH₂)_(n)cycloalkyl, (CH₂)_(n)phenyl, (CH₂)_(n)(substitutedphenyl), (CH₂)_(n)(1 or 2-naphthyl), OR¹⁰, or SR¹¹;

[0102] R¹⁰ and R¹¹ are independently cycloalkyl, phenyl, substitutedphenyl, naphthyl, (CH₂)_(n)cycloalkyl, (CH₂)_(n)phenyl,(CH₂)_(n)(substituted phenyl), or (CH₂)_(n)(1 or 2-naphthyl); and

[0103] n=1-4.

[0104] A fourth group of compounds with respect to the group “A” inFormula I, include those of Formula IIe wherein:

[0105] R⁷ is hydrogen, oxo, cycloalkyl, phenyl, substituted phenyl, ornaphthyl; and

[0106] X=CH₂, (CH₂)₂, (CH₂)₃, or S.

[0107] Another group of compounds with respect to the group “A” inFormula I, include those of Formula IIh wherein:

[0108] a=0 andb=1 or 2.

[0109] Compounds of this invention with respect to the group “B” inFormula I, include those wherein:

[0110] B is hydrogen, 2-benzoxazolyl, substituted 2-oxazolyl, CH₂ZR¹⁵,CH₂O(aryl), or CH₂OPO(R¹⁶)R¹⁷, where Z is an oxygen or a sulfur atom;

[0111] R¹⁵ is phenyl, substituted phenyl, naphthyl, substitutednaphthyl, heteroaryl, (CH₂)_(n)phenyl, (CH₂)_(n)(substituted phenyl),(CH₂)_(n)(1 or 2-naphthyl), or (CH₂)_(n)(heteroaryl);

[0112] R¹⁶ and R¹⁷ are independently alkyl, cycloalkyl, phenyl,substituted phenyl, naphthyl, phenylalkyl, substituted phenylalkyl, or(cycloalkyl)alkyl.

[0113] Another group of compounds with respect to the group “B” inFormula I, include those of Formula IIIa-c wherein:

[0114] Y¹ is O or NR²³;

[0115] Y²is CH₂, O, or NR²³;

[0116] R¹⁸ and R¹⁹ are independently hydrogen, alkyl, or phenyl, or R¹⁸and R¹⁹ taken together are —(CH═CH)₂—;

[0117] R²⁰ is hydrogen, alkyl, phenyl, substituted phenyl,(CH₂)_(n)phenyl, or (CH₂)_(n)(substituted phenyl);

[0118] R²¹, R²² and R²³ are independently hydrogen or alkyl.

[0119] The compounds of Formula I may be synthesized using conventionaltechniques as discussed below. Advantageously, these compounds areconveniently synthesized from readily available starting materials.

[0120] One synthetic route for synthesizing the instant compounds is setforth in the following Scheme 1:

[0121] In the above Scheme 1, Formula (V), that is H₂N—C, is a modifiedaspartic acid residue of Formulas Va through Vd:

[0122] In the above Scheme 1, “PG” stands for an amino protecting groupand “A” stands for a natural or unnatural amino acid of formula Iathrough IIi, as discussed above. In Formula Va through Vd, R²′ is acarboxyl protecting group as described above or an R² moiety as setforth in the definition of R² in Formula I, with the exception that R²′cannot be a hydrogen atom.

[0123] The modified aspartic acids of Formula Va-d can be prepared bymethods well known in the art. See, for example, European PatentApplication 519,748; PCT Patent Application No. PCT/EP92/02472; PCTPatent Application No. PCT/US91/06595; PCT Patent Application No.PCTJUS91/02339; European Patent Application No. 623,592; World PatentApplication No. WO 93/09135; PCT Patent Application No. PCT/US94/08868;European Patent Application No. 623,606; European Patent Application No.618,223; European Patent Application No. 533,226; European PatentApplication No. 528,487; European Patent Application No. 618,233; PCTPatent Application No. PCT/EP92/02472; World Patent Application No. WO93/09135; PCT Patent Application No. PCT/US93/03589; and PCT PatentApplication No. PCT/US93/00481, all of which are herein incorporated byreference.

[0124] The coupling reactions carried out under Step A are performed inthe presence of a standard peptide coupling agent such as thecombination of the combination of dicyclohexylcarbodiimide(DCC) and1-hydroxy-benzotriazole(HOBt), as well as the BOP(benzotriazolyloxy-tris-(dimethylamino)phosphonium hexafluorophosphate)reagent, pyBOP(benzotriazolyloxy-tris(N-pyrolidinyl)phosphoniumhexafluoroplospbate),TU (O-benzotriazolyly-tetramethylisouronium-hexafluorophosphate), andEEDQ (1-ethyloxycarbonyl-2-ethyloxy-1,2-dihydroquinoline) reagents, thecombination of 1-ethyl(3,3′-dimethyl-1′-aminopropyl)carbodiimide (EDAC)and HOBt, and the like, as discussed in J. Jones, “Amino Acid andPeptide Synthesis,” Steven G. Davis ed., Oxford University Press,Oxford, pp. 25-41 (1992); M. Bodanzky, “Principles of PeptideSynthesis,” Hafner et al. ed., Springer-Verlag, Berlin Heidelberg, pp.9-52 and pp. 202-251 (1984); M. Bodanzky, “Peptide Chemistry, APractical Textbook,” Springer-Verlag, Berlin Heidelberg, pp. 55-73 andpp. 129-180; and Stewart and Young, “Solid Phase Peptide Synthesis,”Pierce Chemical Company, (1984), all of which are herein incorporated byreference. The amino protecting group is then removed and the resultingamine is coupled to the (N-substituted) oxamic acid of Formula VII (StepB). Again, this coupling reaction uses the standard peptide couplingreactions mentioned above.

[0125] Alternatively, the (N-substituted)oxamic acid of Formula VII canbe coupled to an amino ester of Formula IX (Step D). Again, thiscoupling reaction uses the standard peptide coupling reactions mentionedabove. In Formula IX, the group R is a carboxyl protecting group such asmethyl, allyl, benzyl or tert-butyl. After removal of the carboxylprotecting group under standard conditions well known in the art, theresulting carboxylic acid is coupled to amine V using the standardpeptide coupling methods described above (Step E).

[0126] In the case where the coupling reaction depicted by either Step Aor Step E was carried out with the amino alcohol of Formula Vc, thealcohol moiety must be oxidized to the corresponding carbonyl compoundprior to removal of the protecting groups. Preferred methods for theoxidation reaction include Swern oxidation (oxalyl chloride-dimethylsulfoxide, methylene chloride at −78° C. followed by triethylamine); andDess-Martin oxidation (Dess-Martin periodinane, t-butanol, and methylenechloride.) The protecting groups contained in substructures of theFormula Va-d and AL (if present) are removed by methods well known inthe art. These reactions and removal of some or all of the protectinggroups are involved in Step C in the above Scheme 1.

[0127] An alternative synthetic route for synthesizing the instantcompounds is set forth in the following Scheme 2:

[0128] In the above Scheme 2, “PG” stands for an amino protecting groupand “A” stands for a natural or unnatural amino acid of formula IIathrough IIi, as discussed above. The group R is a carboxyl protectinggroup such as trimethylsilyl, methyl, allyl, benzyl or tert-butyl.

[0129] The coupling reactions carried out under Step F and Step G areperformed in the presence of a standard peptide coupling agent asdiscussed above. In Step Q the amino protecting group must be removedprior to the coupling step. In Step H the alpha-carboxy protecting groupR of the compound of Formula XIII is selectively removed and theresulting mono-carboxylic acid treated sequentially with diazomethaneand hydrobromic acid to give the alpha-bromoketone of Formula XIV InStep I, the bromoketone of Formula XIV is treated with either R¹⁵Z—H,(aryl)-CO₂H, (heteroaryl)-CO₂H, or R¹⁶(R¹⁷)PO₂H in the presence of aninorganic base such as potassium carbonate or potassium fluoride in aninert solvent such as dimethyl formamide to give the correspondingcompound of Formula I in which B is CH₂ZR¹⁵, CH₂OCO(aryl),CH₂OCO(heteroaryl), or CH₂OPO(R¹⁶)R¹⁷, respectively. Compounds ofFormula I in which B is a fragment of Formula III may also be preparedin a similar fashion. The protecting groups contained in substructuresof the Formula XI and A are removed by methods well known in the art.These reactions and removal of some or all of the protecting groups areinvolved in Step I in the above Scheme 2.

[0130] An alternative method for the preparation of compounds of theinstant invention of Formula I in which R² and B are both hydrogen(i.e., Formula Ib) is set forth below in Scheme 3:

[0131] In Scheme 3, Fmoc is the amino protecting group9-fluorenylmethoxycarbonyl and the shaded circle labeled “PS” representspolystryene resin.

[0132] The coupling of the acid of Formula XV to a primary amine onsolid support, preferably aminomethyl polystyrene, is carried out usingstandard peptide coupling agents, preferably usingbenzotriazolyloxy-tris(N-pyrolidinyl)phosphoniumhexafluorophosphate(pyBOP) in a inert solvent such as dimethylformamide or N-methylpyrrolidone (Step I). After removal of the Fmoc protecting group of XVIby treatment with pyrrolidine-dimethylformamide, the resulting amine iscoupled to Fmoc-amino acid of Formula IVa using standard peptidecoupling conditions as discussed above (Step K).

[0133] In Step L the Fmoc protecting group of the compound of FormulaXVII is removed again by treatment with pyrrolidine-dimethylformamideand the resulting amine coupled to the (N-substituted)oxamic acid ofFormula VII again using standard peptide coupling conditions asdiscussed above. The tert-butyl ester of the compound of Formula XVIIIis removed by treatment with trifluoroacetic acid-methylene chloride inthe presence of a trapping agent such as anisole and the resulting acidcleaved from the solid support by treatment with 37% aqueousformaldehyde/acetic acid/tetrahydrofuran/trifluoroacetic acid,preferably in a ratio of 1/1/5/0.025, to give the aspartyl aldehyde ofFormula lb (Step M).

[0134] Pharmaceutical compositions of this invention comprise any of thecompounds of the present invention, and pharmaceutically acceptablesalts thereof, with any pharmaceutically acceptable carrier, adjuvant orvehicle (hereinafter collectively referred to as“pharmaceutically-acceptable carriers”). Pharmaceutically acceptablecarriers, adjuvants and vehicles that may be used in the pharmaceuticalcompositions of this invention include, but are not limited to, ionexchange, alumina, aluminum stearate, lecithin, serum proteins, such ashuman serum albumin; buffer substances such as the various phosphates,glycine, sorbic acid, potassium sorbate, partial glyceride mixtures ofsaturated vegetable fatty acids; water, salts or electrolytes, such asprotamine sulfate, disodium hydrogen phosphate, potassium hydrogenphosphate, sodium chloride, and zinc salts; colloidal silica, magnesiumtrisilicate, polyvinyl pyrrolidone, cellulose-based substances,polyethylene glycol, sodium carboxymethylcellulo se, polyarylates,waxes, polyethylene-polyoxypropylene-block polymers, polyethylene glycoland wool fat, and the like.

[0135] The pharmaceutical compositions of this invention may beadministered orally, parenterally, by inhalation spray, topically,rectally, nasally, buccally, vaginally or by an implanted reservoir.Oral and parenteral administration are preferred. The term “parenteral”as used herein includes subcutaneous, intracutaneous, intravenous,intramuscular, intra-articular, intrasynovial, intrasternal,intrathecal, intralesional and intracranial injection or infusiontechniques.

[0136] The pharmaceutical compositions may be in the form of a sterileinjectable preparation, for example, as a sterile injectable aqueous oroleaginous suspension. This suspension may be formulated according totechniques known in the art using suitable dispersing or wetting agents(such as, for example, Tween 80) and suspending agents. The sterileinjectable preparation may also be a sterile injectable solution orsuspension in a non-toxic parenterally-acceptable diluent or solvent,for example, as a solution in 1,3-butanediol. Among the acceptablevehicles and solvents that may be employed are mannitol, water, Ringer'ssolution and isotonic sodium chloride solution. In addition, sterile,fixed oils are conventionally employed as a solvent or suspendingmedium. For this purpose, any bland fixed oil may be employed includingsynthetic mono- or diglycerides. Fatty acids, such as oleic acid and itsglyceride derivatives are useful in the preparation of injectables, asare natural pharmaceutically-acceptable oils, such as olive oil orcastor oil, especially in their polyoxyethylated versions. These oilsolutions or suspensions may also contain a long-chain alcohol diluentor dispersant.

[0137] The pharmaceutical compositions of this invention may be orallyadministered in any orally acceptable dosage form including, but notlimited to, capsules, tablets, and aqueous suspensions and solutions. Inthe case of tablets for oral use, carrier which are commonly usedinclude lactose and corn starch. Lubricating agents, such as magnesiumstearate, are also typically added. For oral administration in capsuleform useful diluents include lactose and dried corn starch. When aqueoussuspensions are administered orally, the active ingredient is combinedwith emulsifying and suspending agents. If desired, certain sweeteningand/or flavoring and/or coloring agents may be added.

[0138] The pharmaceutical compositions of this invention may also beadministered in the form of suppositories for rectal administration.These compositions can be prepared by mixing a compound of thisinvention with a suitable non-irritating excipient which is solid atroom temperature but liquid at the rectal temperature. Such materialsinclude, but are not limited to, cocoa butter, beeswax and polyethyleneglycols.

[0139] Topical administration of the pharmaceutical compositions of thisinvention is especially useful when the desired treatment involves areasor organs readily accessible to topical application. For applicationtopically to the skin, the pharmaceutical composition should beformulated with a suitable ointment containing the active componentssuspended or dissolved in a carrier. Carriers for topical administrationof the compounds of this invention include, but are not limited to,mineral oil, liquid petroleum, white petroleum, propylene glycol,polyoxyethylene, polyoxypropylene compound, emulsifying wax and water.Alternatively, the pharmaceutical composition can be formulated with asuitable lotion or cream containing the active compound suspended ordissolved in a carrier. Suitable carriers include, but are not limitedto, mineral oil, sorbitan monostearate, polysorbate 60, cetyl esterswax, cetearyl alcohol, 2-octyldodecanol, benzyl alcohol and water. Thepharmaceutical compositions of this invention may also be topicallyapplied to the lower intestinal tract by rectal suppository formulationor in a suitable enema formulation. Topically-applied transdermalpatches are also included in this invention.

[0140] The pharmaceutical compositions of this invention may beadministered by nasal aerosol or inhalation. Such compositions areprepared according to techniques well-known in the art of pharmaceuticalformulation and may be prepared as solutions in saline, employing benzylalcohol or other suitable preservatives, absorption promoters to enhancebioavailability, fluorocarbons, and/or other solubilizing or dispersingagents known in the art.

[0141] The compounds of this invention may be used in combination witheither conventional anti-inflammatory agents or with matrixmetalloprotease inhibitors, lipoxygenase inhibitors and antagonists ofcytokines other than IL-1β.

[0142] The compounds of this invention can also be administered incombination with immunomodulators (e.g., bropirimine, anti-human alphainterferon antibody, IL-2, GM-CSF, methionine enkephalin, interferonalpha, diethyldithiocarbamate, tumor necrosis factor, naltrexons andrEPO) or with prostaglandins, to prevent or combat IL-1-mediated diseasesymptoms such as inflammation.

[0143] When the compounds of this invention are administered incombination therapies with other agents, they may be administeredsequentially or concurrently to the patient. Alternatively,pharmaceutical compositions according to this invention may be comprisedof a combination of a compound of Formula I and another therapeutic orprophylactic agent mentioned above.

[0144] The disease states which may be treated or prevented by theinstant pharmaceutical compositions include, but are not limited to,inflammatory diseases, autoimmune diseases and neurodegenerativediseases, and for inhibiting unwanted apoptosis involved in ischemicinjury, such as ischemic injury to the heart (e.g., myocardialinfarction), brain (e.g., stroke), and kidney (e.g., ischemic kidneydisease). As a consequence of their ability to inhibit apoptosis, thepresent pharmaceutical compositions are also useful for the repopulationof hematopoietic cells of a patient following chemotherapy. Methods ofadministering an effective amount of the above-described pharmaceuticalcompositions to mammals, also referred to herein as patients, in need ofsuch treatment (that is, those suffering from inflammatory diseases,autoimmune diseases, neurodegenerative diseases and for the repopulationof hematopoielic cells in cancer patients who have undergonechemotherapy) are another aspect of the instant invention. Finally, as afurther consequence of their ability to inhibit apoptosis, the instantpharmaceutical compositions may be used in a method to prolong theviability of organs to be used in transplantations.

[0145] Inflammatory disease which may be treated or prevented include,for example, septic shock, septicemia, and adult respiratory distresssyndrome. Target autoimmune diseases include, for example, rheumatoid,arthritis, systemic lupus erythematosus, scleroderma, chronicthyroiditis, Graves' disease, autoimmune gastritis, insulin-dependentdiabetes mellitus, autoimmune hemolytic anemia, autoimmune neutropenia,thrombocytopenia, chronic active hepatitis, myasthenia gravis andmultiple sclerosis. Target neurodegenerative diseases include, forexample, amyotrophic lateral sclerosis, Alzheimer's disease, Parkinson'sdisease, and primary lateral sclerosis. The pharmaceutical compositionsof this invention may also be used to promote wound healing. Targetdiseases associated with harmful, apoptosis, in other words, thoseassociated with ischemic injury, includes myocardial infarction, stroke,and ischemic kidney disease. The pharmaceutical compositions of thisinvention may also be used to treat infectious diseases, especiallythose involved with viral infections.

[0146] The term “effective amount” refers to dosage levels of the orderof from about 0.05 milligrams to about 140 milligrams per kilogram ofbody weight per day for use in the treatment of the above-indicatedconditions (typically about 2.5 milligrams to about 7 grams per patientper day). For example, inflammation may be effectively treated by theadministration of from about 0.01 to 50 milligrams of the compound perkilogram of body weight per day (about 0.5 milligrams to about 3.5 gramsper patient per day).

[0147] The amount of the compounds of Formula I that may be combinedwith the carrier materials to produce a single dosage form will varydepending upon the host treated and the particular mode ofadministration. For example, a formulation intended for the oraladministration of humans may contain from 0.5 milligrams to 5 grams of acompound of Formula I combined with an appropriate and convenient amountof a pharmaceutically-acceptable carrier which may vary from about 5 toabout 95 percent of the total composition. Dosage unit forms willgenerally contain between from about 1 milligram to about 500 milligramsof an active compound of Formula I.

[0148] It will be understood, however, that the specific “effectiveamount” for any particular patient will depend upon a variety of factorsincluding the activity of the specific compound employed, the age, bodyweight, general health, sex, diet, time of administration, route ofadministration, rate of excretion, drug combination and the severity ofthe particular disease undergoing prevention or therapy.

[0149] Although this invention focuses on the use of the compoundsdisclosed herein for preventing and treating IL-1-mediated diseases, thecompounds of this invention can also be used as inhibitory agents forother cysteine proteases.

[0150] The compounds of this invention are also useful as commercialreagents which effectively bind to the ICE/ced-3 family of cysteineprotease or other cysteine proteases. As commercial reagents, thecompounds of this invention, and their derivatives, may be used to blockproteolysis of a target peptide or may be derivatized to bind to astable resin as a tethered substrate for affmity chromatographyapplications. These and other uses which characterize commercialcysteine protease inhibitors will be evident to those of ordinary skillin the art.

[0151] In order that this invention be more fully understood, thefollowing examples are set forth. These examples are for the purpose ofillustration only and are not to be construed as limiting the scope ofthe invention in any way. In the following Examples, proton NMR spectrawere obtained at 300 MHz; chemical shifts are quoted downfield frominternal tetramethylsilane.

PREPARATION 1

[0152]

Preparation of (3 S)-Amino-4-Oxobutanoic Acid (Tert)-Butyl EsterSemicarbazone, P-Toluenesulfonate Salt

[0153] Part A: N-(Benzyloxycarbonyl)-L-(N′-Methyl-N′-Methoxy)aspartamideD-(tert-Butyl) Ester

[0154] To a solution of N-(benzyloxycarbonyl)-L-asparticacid-β-(tert-butyl) ester (14.65 g, 45.3 mmol, Bachem) in CH₂Cl₂ (150mL) at 0° C. (ice bath) under a nitrogen atmosphere was added1-hydroxybenzotriazole hydrate (7.29 g, 47.6 mmol, Aldrich) followed by1-ethyl-3-(3′,3′-dimethyl-1′-aminopropyl)carbodiimide hydrochloride(9.55 g, 49.8 mmol, Sigma). After stirring at 0° C. for 15 min.,N,O-dimetbylhydroxylamine hydrochloride (5.10 g, 52.3 mmol, Aldrich) andN-methylmorpholine (5.8 mL, 53 mmol, Aldrich) were added. The mixturewas allowed to warm to room temperature over 3 hours then stirred atroom temperature for 16 hours. The solution was concentrated undervacuum and the residue partitioned between ethyl acetate-5% KHSO₄ (200mL each). The organic phase was washed in turn with 5% KHSO4, saturatedsodium bicarbonate and saturated sodium chloride solutions; dried overanhydrous sodium sulfate and evaporated to an oil. The oil wascrystallized from hexane to give the title product (16.10 g, 97% yield)as a fluffy white crystalline solid. TLC (ethyl acetate), single spot(UV and PMA): Rf=0.37.

[0155] A similar procedure to the one above, starting with 29.3 g ofN-(benzyloxycarbonyl)-L-aspartic acid-o-(tert-butyl)ester (2-fold scaleup) gave 31.18 g (94% yield) of the title product.

[0156] Part B: (3S)-(L3enzyloxycarbonyl)Amino-4-Oxobutanoic Acid(tert)-Butyl Ester Semicarbazone.

[0157] To a solution ofN-(benzyloxycarbonyl)-L-(N′-methyl-N′-methoxy)aspartamide-O-(tert-butyl)ester (15.50 g, 42.3 mmol) in anhydrous ether (400 mL) at 0+ C. (icebath) under a nitrogen atmosphere was added dropwise to a 1.0 M solutionof LiA1H4 in ether (22.0 mL, 22.0 mmol, Aldrich) at such a rate as tokeep the reaction solution temperature between 0-5° C. (addition time15-20 min). After the addition of the lithium aluminum hydride reagentwas complete, the mixture was stirred at 0-5° C. for 1 hr, then quenchedby the dropwise addition of 0.3 N KHSO₄ solution (100 mL). The resultantmixture was transferred to a separatory funnel adding sufficient 5%KHSO₄ solution (75 mL) to dissolve the solids. The organic phase wasseparated and the combined aqueous washes back-extracted with ether (100mL). The combined ether extracts were washed with saturated NaClsolution, dried over anhydrous sodium sulfate and concentrated in vacuowith minimal heating. TLC (ethyl acetate): streaky spot (UV and PMA)Rf=0.48. TLC (methanol/methylene chloride, 1:9) major spot (UV and PMA):Rf=0.75.

[0158] The crude aldehyde was immediately taken up in aqueous ethanol(45 mL water/105 mL alcohol), placed in an ice bath and treated withsodium acetate (3.82 g, 46.6 mmol) and semicarbazide hydrochloride (5.20g, 46.6 mmol, Aldrich). The mixture was stirred at 0° C. (ice bath)under a nitrogen atmosphere for 3 hrs, allowed to warm to roomtemperature, and stirred overnight (16 hrs). Most of the ethanol wasremoved under vacuum and the residue partitioned between ethyl acetateand water (100 mL each). The organic phase was washed sequentially with5% KHSO₄, saturated sodium bicarbonate and saturated sodium chloridesolutions; dried over anhydrous sodium sulfate and evaporated todryness. The crude product of this reaction was combined with that oftwo similar procedures starting with 15.40 g and 4.625 g ofN-(benzyloxycarbonyl)-L-(N′-methyl-N-methoxy)aspartamide-O-(tert-butylester) (total: 35.525 g, 97 mmol) and these combined products werepurified by flash chromotagraphy on silica gel eluting withacetone/methylene chloride (3:7) then methanol-acetone-methylenechloride (0.5:3:7) to give pure title product (27.73 g, 78.5%) as acolorless foam. TLC (MeOH—CH₂Cl₂, 1:9): single spot (UV and PMA),Rf=0.51.

[0159] Part C: (3S)-Amino-4-Oxobutanoic Acid (tert)-Butyl EsterSemicarbazone. p-Toluenesulfonate Salt

[0160] To a solution of (3S)-(benzyloxycarbonyl)amino-4-oxobutanoic acid(tert)-butyl ester semicarbazone (13.84 g, 38.0 mmol) in absoluteethanol (250 mL) was added 10% Pd/C (1.50 g, Aldrich) and the resultingmixture stirred under an atmosphere of hydrogen (balloon) until TLC(methanol/methylene chloride, 1:9) indicated complete consumption of thestarting material (60 min). Note: It is important to follow thisreaction closely since the product can be over-reduced. The mixture wasfiltered though Celite and evaporated to an oil. The oil was chased withmethylene chloride (2×75mL) then with methylene chloride/toluene (1:1,75 mL) to give the crude amine as a white crystalline solid. TLC(EtOAc-pyridine-AcOH—H₂O; 60:20:5:10) single spot (UV and PMA) Rf=0.24.Note: In this TLC system, any over-reduced product will show upimmediately below the desired product, Rf=0.18 (PMA only).

[0161] The crude amine was taken up in CH₃CN (60 mL) and treated with asolution of p-toluenesulfonic acid monohydrate (7.22 g, 38.0 mmol) inacetonitrile (60 mL). The crystalline precipitate was collected, washedwith acetonitrile and ether, and air-dried to give the title compound(13.95 g, 92% yield) as a white, crystalline solid.

[0162] The optical purity of this material was checked by conversion tothe corresponding Mosher amide [1.05 equiv(R)-(−)-α-methoxy-α-(trifluoromethyl)phenylacetyl chloride, 2.1equivalents of i-Pr₂NEt in CH₂Cl₂, room temperature, 30 min]. Thedesired product has a doublet at 7.13 ppm (1H, d, J=2.4 Hz, CH═N) whilethe corresponding signal for its diastereomer is at 7.07 ppm. Theoptical purity of the title compound obtained from the above procedureis typically >95:5.

PREPARATION 2

[0163]

Preparation of (3 S)-(9-Fluorenylmethoxycaonyl)Amino-4-Oxobutanoic Acid(Tert)-Butyl Ester Semicarbazonyl4-[2′-(4-Ethyl-Phenoxyacetic Acid)]

[0164] Part A: 4-[2′-(N-t-Butoxycarbonyl)Aminoethyl]Phenoxyacetic Acid,Methyl Ester

[0165] To a suspension 4-hydroxy-phenethylanune (7.00 g, 51.1 mmol,Aldrich) in dry dimethylformamide (50 mL) at room temperature undernitrogen was added di-tert-butyl dicarbonate (11.0 g, 50.5 mmol). Afterstirring at room temperature for 1 hr; the resulting clear solution wastreated with methyl bromoacetate (7.5 mL, 79 mmol) and cesium carbonate(17.5 g, 53.7 mmol). After stirring at room temperature for 16 hrs, TLC(Et₂O-toluene; 2:8) shows some unalkylated material remained (Rf=0.43)and a second portion of methyl bromoacetate (2.0 mL, 21 mmol) and cesiumcarbonate (4.5 g, 14 mmol) were added. After stirring for an additional24 hrs, the mixture was partitioned between EtOAc-water (250 mL each),organic phase washed successively with water (3X), 5% potassiumbisulfate and saturated NaCl solutions, dried over anhydrous Na₂SO₄ andevaporated to dryness. Trituration of the residue with hexane gave 15.87g of a tan solid. Filtration of the crude product through a pad ofsilica gel eluting with EtOAc-hexane (2:8) and crystallization fromhexane gave the title compound (14.75, 93%) as a white granular,crystalline solid. TLC (Et₂O-toluene; 2:8) Rf=0.53.

[0166] Part B: 4-(2′-Aminoethyl)Phenoxyacetic Acid, Methyl Ester,Hydrochloride

[0167] To a solution 4-[2′-(N-t-butoxycarbonyl) aminoethyl]phenoxyaceticacid, methyl ester (18.31 g, 59.3 mmol) in dioxane (55 mL) at roomtemperature was added 4.0 N HCl in dioxane (55 mL). After stirring atroom temperature for 16 hrs, the mixture was diluted with Et₂O, theprecipitate collected, washed thoroughly with Et₂O and dried in vacuo togive the title compound (14.55 g, 94%) was a fluffy white, crystallinesolid.

[0168] Part C:1-tert-Butoxycarbonyl-Semicarbazidyl-4-[2′-(4-Ethyl-Phenoxyacetic Acid)]Methyl Ester.

[0169] A solution of t-butyl carbazate (6.60 g, 50 mmol) indimethylformamide (50 mL) was added dropwise to a solutioncarbonyldiimidazole (8.10 g, 50 mmol) in dimethylformamide (80 niL) over40 min at room temperature under nitrogen. After stirring at roomtemperature for an additional 30 min, 4-(2′-aminoethyl)phenoxyaceticacid, methyl ester, hydrochloride (12.3 g, 50 mmol) was added as a solidin one portion followed by a triethylamine (8.0 niL, 58 mmol) addeddropwise over 30 min. After stirring at room temperature for 18 hrs, themixture was partitioned between EtOAc-water (300 niL each). The organicphase was washed successively with water (3X), 5% potassium bisulfate,saturated sodium bicarbonate, and saturated NaCl solutions, dried overanhydrous Na₂SO₄ and evaporated to dryness. Crystallization of theresidue from EtOAc-hexane gave the title compound (15.50, 84%) as anoff-white crystalline solid. TLC (MeOH-CH₂Cl₂; 1:9) Rf=0.45.

[0170] Part D: 1-tert-Butoxcarbonyl-Semicarbazidyl-4-[2′-(4-Ethyl-Phenoxyacetic Acid)]

[0171] A solution of1-tert-butoxycarbonyl-semicarbazidyl-4-[2′-(4-ethyl-phenoxyacetic acid)]methyl ester (14.68 g, 40 mmol) in dioxane (50 niL) at room temperatureunder nitrogen was added 1.0 N LiOH solution (50 mL). After stirring atroom temperature for 1 hr, the mixture was acidified with conc. HCl andextracted with EtOAc (100 mL). The organic phase was washed withsaturated NaCl solution, dried over anhydrous Na₂SO₄ and evaporated to awhite solid. Recrystallization of the crude product fromTHF-EtOAc-hexane gave the title compound (13.44, 95%) as a whitecrystalline solid. TLC (AcOH—MeOH—CH₂Cl₂; 1:1:8) Rf=0.31.

[0172] Part E: Semicarbazidyl-4-[2′-(4-Ethyl-Phenoxyacetic Acid)]Hydrochloride

[0173] To a solution of1-tert-butoxycarbonyl-senicarbazidyl-4-[2′-(4-ethyl-phenoxyacetic acid)](13.43 g, 38.0 mmol) in dioxane (80 mL)-anisole (15 mL) at roomtemperature was added 4.0 N HCl in dioxane (35 mL). After stirring atroom temperature for 18 hrs, additional 4.0 N HCl in dioxane (15 mL) wasadded. After an additional 6 hrs, the precipitate was collected, washedthoroughly with dioxane then Et₂O and dried in vacuo to give the titlecompound (11.67 g, 100%) was a white, crystalline solid.

[0174] Part F:N-(9-Fluorenylmethoxycarbonyl)-L-(N′-Methyl-N′-Methoxyaspartamideβ-(tert-Butyl) Ester

[0175] To a solution of N-(9-fluorenylmethoxycarbonyl)-L-asparticacid-β-(tert-butyl) ester (16.48 g, 40 mmol) in CH₂Cl₂ (80mL)-tetrahydrofuran (20 mL) at 0° C. (ice bath) under a nitrogenatmosphere was added 1-hydroxybenzotriazole hydrate (7.12 g, 46.5 mmol)followed by 1-ethyl-3-(3′,3′-dimethyl-1′-aminopropyl)carbodiimidehydrochloride (9.20 g, 48 mmol). After stirring at 0° C. for 15 min.,N,O-dimethylhydroxylamine hydrochloride (4.68 g, 48 mmol) andN-methylmorpholine (5.2 mL, 47 mmol) were added. The mixture was allowedto warm to room temperature over 2 hours then stirred at roomtemperature for 16 hours. The solution was concentrated under vacuum andthe residue partitioned between ethyl acetate-5% KHSO₄ (200 mL each).The organic phase was washed successively with 5% KHSO₄, saturatedsodium bicarbonate and saturated sodium chloride solutions; dried overanhydrous sodium sulfate and evaporated to an oil. Purification of thecrude product by flash chromatography on silica gel eauting withEtOAc-hexane (30:70 then 35:65) gave the title product (17.75 g, 98%yield) as a colorless foam. TLC (EtOAc-hexane; 1:1) Rf=0.35.

[0176] Part G: (3 S)-(9-Fluorenylmethoxycabonyl)Amino-4-Oxobutanoic Acid(tert)-Butyl Ester Semicarbazonyl-4-[2′-(4-Ethyl-Phenoxyacetic Acid)]

[0177] To a solution ofN-(9-fluorenylmethoxycarbonyl)-L-(N′-methyl-N′-methoxy)aspartamide-β-(tert-butyl)ester (13.20 g, 29 mmol) in anhydrous ether (250 mL) at 0° C. (ice bath)under a nitrogen atmosphere was added dropwise to a 1.0 M solution ofLiAIH₄ in ether (14.5 mL, 14.5 mmol) at such a rate as to keep thereaction solution temperature between 0-5° C. (addition time 15-20 min).After the addition of the lithium aluminum hydride reagent was complete,the mixture was stirred at 0-5° C. for 1 hr. then quenched by thedropwise addition of 0.3 N KHSO₄ solution (100 mL). After addingsufficient 0.3 N KHSO₄ solution to dissolve most of the inorganic salts,the mixture was transferred to a separatory funnel. The organic phasewas separated and the aqueous phase back-extracted with ether (100 mL).The combined ether extracts were washed with saturated NaCl solution,dried over anhydrous sodium sulfate and concentrated in vacuo withminimal heating. TLC (EtOAc-bexane): Rf=0.40.

[0178] The crude aldehyde was immediately taken up in ethanol(105mL)-water(45 mL)-tetrahydrofuran(75 mL), placed in an ice bath andtreated with sodium acetate (3.20 g, 39 mmol) andsemicarbazidyl-4-[2′-(4-ethyl-phenoxyacetic acid)] hydrochloride (8.65g, 30 mmol). The mixture was stirred at 0+ C. (ice bath) under anitrogen atmosphere for 3 hrs, allowed to warm to room temperature, andstirred overnight (16 hrs). The mixture was concentrated on a rotovap,diluted with water and resulting precipitate collected by suction. Thematerial was dried in vacuo to give 18.36 g of crude product as a whitesolid. The crude product of this reaction was combined with that of asmaller scale reaction (6.34 g) starting with 4.55 g (10 mmol) ofN-(9-fluorenylmethoxycarbonyl)-L-(N′-methyl-N′-methoxy)aspartamide-β-(tert-butylester) and partitioned between ethyl acetate-tetrahydrofuran(1:1) and 5%KHSO₄. The organic phase was washed with 5% KHSO₄ and saturated sodiumchloride solutions, dried over anhydrous sodium sulfate and evaporatedto dryness. The residue was purified by filtration through a pad ofsilica gel eluting with terahydrofuran/methylene chloride (1:1). Thecombined product-containing fractions were evaporated to dryness andrecrystallized from tetrahydrofuran-Et₂O to give pure title product(17.01 g, 69%) as awhite solid. TLC (AcOH—MeOH—CH₂Cl₂, 1:1:40): Rf=0.19.

PREPARATION 3 Assay for Inhibition of ICE/ced-3 Protease Family Activity

[0179] A. Deterrnination of IC₅₀ Values

[0180] Fluorescence enzyme assays detecting the activity of thecompounds of Formula I utilizing the recombinant ICE and CPP32 enzymesare performed essentially according to Thomberry et al. (Nature,356:768:774 (1992)) and Nicholson et al. (Nature, 376:37-43 (1995))respectively, (herein incorporated by reference) in 96 well microtiterplates. The substrate is Acetyl—Tyr—Val—Ala—Asp—amino-4-methylcoumarin(AMC) for the ICE assay andAcetyl—Asp—Glu—Val—Asp—amino-4-methylcoumarin for the CPP32, Mch2, Mch3and Mch5 assays. Enzyme reactions are run in ICE buffer (25 mM HEPES, 1mM EDTA, 0.1% CHAPS, 10% sucrose, pH 7.5) containing 2 mM DTT at roomtemperature in duplicate. The assays are performed by mixing thefollowing components:

[0181] 50 μL ICE, Mch2, Mch5, CPP32 (18.8, 38, 8.1 and 0.153 nMconcentrations, respectively) or Mch3 (1 unit) enzyme in ICE buffercontaining either 8.0 (ICE, Mch2, Mch3, CPP32) or 20 (Mch5) mM DTT;

[0182] 50 μL compound of Formula I or ICE buffer (control); and

[0183] 100 μL of 20 μM substrate.

[0184] The enzyme and the compound of Formula I to be assayed areallowed to preincubate in the microtitre plate wells for 30 minutes atroom temperature prior to the addition of substrate to initiate thereaction. Fluorescent AMC product formation is monitored for one hour atroom temperature by measuring the fluorescence emission at 460 nm usingan excitation wavelength of 360 nm. The fluorescence change in duplicate(control) wells are averaged and the mean values are plotted as afuinction of inhibitor concentration to determine the inhibitorconcentration producing 50% inhibition (IC₅₀).

[0185] B. Determination of the dissociation constant Ki and irreversiblerate constant k₃ for irreversible inhibitors

[0186] For the irreversible inhibition of a ICE/ced-3 Family Proteaseenzyme with a competitive irreversible inhibitor; using the modelrepresented by the following formulas:

[0187] The product formation at time t may be expressed as:$\begin{matrix}{\lbrack P\rbrack_{t} = {\lbrack E\rbrack^{T}( \frac{\lbrack S\rbrack K_{i}}{\lbrack I\rbrack K_{s}} ){( \frac{k_{s}}{k_{3}} )\lbrack {1 - ^{{- k_{3}}{t/{({1 + {\frac{K_{i}}{\lbrack I\rbrack}{({1 + \frac{\lbrack S\rbrack}{K_{s}}})}}})}}}} \rbrack}}} & {{Equation}\quad 1}\end{matrix}$

[0188] where E, I, El and E-I denote the active enzyme, inhibitor,non-covalent enzyme-inhibitor complex and covalent enzyme-inhibitoradduct, respectively. The K_(i) value is the overall dissociationconstant of the reversible binding steps, and k₃ is the irreversiblerate constant. The [S] and K_(S) values are the substate concentrationand dissociation constant of the substrate bound to the enzyme,respectively. [E]^(T) is the total enzyme concentration.

[0189] The above equations are used to determine the Ki and k₃ values ofa given inhibitor bound to a ICE/ced-3 family protease. Thus, acontinuous assay is run for sixty minutes at various concentrations ofthe inhibitor and the substrate. The assay is formulated essentially thesame as described above for generating the data in Table 1, except thatthe reaction is initiated by adding the enzyme to thesubstrate-inhibitor mixture. The K_(i) and k₃ values are obtained bysimulating the product AMC formation as a function of time according toEquation 1.

[0190] The following are examples of compounds of the invention.

EXAMPLE 1

[0191]

[0192] (3 S)-3-[N-(N′-( 1-NaphythylOxamylLeucinyl]Amino-4-OxobutanoicAcid

[0193] Part A: N-(1-Naphthyl)Oxamic Acid

[0194] To a solution of I-aminonaphthylene (1.43 g, 10 mmol) andtriethylamine (1.5 mL, 10.8 mmol) in CH₂Cl₂ (10 mL) at 0° C. (ice bath)under nitrogen was added dropwise a solution of methyl oxalyl chloride(1.0 mL, 10.9 mmol) in CH₂Cl₂ (5 mL). When the addition was complete,the mixture was allowed to come to room temperature and stirred for 1hr. The mixture was concentrated and the residue partitioned betweenEtOAc-5% KHSO₄. The organic phase was washed with 5% KHSO₄ and saturatedNaCl solutions, dried over anhydrous Na₂SO₄ and evaporated to a pinksolid. Recrystallization of the crude product from toluene-hexane gavethe N-(1-naphthyl)oxamic acid methyl ester (2.066 g, 90%) as a pinkcrystalline solid. TLC(EtOAc-hexane) Rf=0.6.

[0195] The methyl ester (1.97 g, 8.6 mmol) was taken up in dioxane (10mL) and treated with 1.0 N LiOH solution (10 mL, 10 mmol) and stirred atroom temperature for 1 hr. The mixture was acidified with conc. HCl andextracted with EtOAc. The extract was washed with saturated NaClsolution, dried over anhydrous Na₂SO₄ and evaporated to a pink solid.Recrystallization of the crude product from EtOAc-hexane gave the titlecompound (1.712 g, 85%) as a pink crystalline solid.TLC(AcOH—MeOH—CH₂Cl₂; 1:1:20) Rf=0.06.

[0196] Part B: (3S)-3-[(N-Benzyloxycarbonyl)Leucinyl]Amino-4-OxobutanoicAcid (tert)-Butyl Ester Semicarbazone

[0197] To a solution of (N-benzyloxycarbonyl)leucine N-hydroxysuccinimdeester (1.81 g, 5.0 mmol) in CH₂Cl₂ (30 mL) at room temperature undernitrogen was added (3S)-amino-4-oxobutanoic acid (tert)-butyl estersemicarbazone, p-toluenesulfonate salt (2.58 g, 6.4 mmol) followed bydiisopropyl ethylamine (1.2 mL, 6.9 mmol). After stirring at roomtemperature for 16 hrs, the mixture was concentrated and the residuepartitioned between EtOAc-5% KHSO₄. The organic phase was washed with 5%KHSO₄, saturated NaHCO₃ and saturated NaCl solutions, dried overanhydrous Na₂SO₄ and evaporated to give the title compound (2.798 g) asa pale yellow foam. TLC(MeOH—CH₂Cl₂; 1:9) Rf=0.52.

[0198] Part C: (3 S)-3-(Leucinyl)Amino-4-Oxobutanoic Acid (tert)-ButylEster Semicarbazone

[0199] To a solution of crude (3S)-[(N-benzyloxycarbonyl)leucinyl]amino-4-oxobutanoic acid (tert)-butylester semicarbazone (2.798 g, ca.5.0 mmol) in absolute EtOH (40 mL) wasadded 10% Pd-C (0.40 g) and resulting mixture stirred under a hydrogenatmosphere (balloon) for 1.5 hrs. The mixture was filtered throughCelite washing the filter cake with CH₂Cl₂ and the combined filtratesevaporated to dryness. The residue was chased with CH₂Cl₂ (2×20 mL) togive the title product (2.113 g) as a colorless foam. TLC(MeOH—CH₂Cl₂;1:9) Rf=0.23.

[0200] Part D:(3S)-3-[N-(N′-(1-Naphthyl)Oxamyl)Leucinyl]Amino-4-Oxobutanoic Acid(tert)-Butyl Ester Semicarbazone

[0201] To a solution of N-(1-naphthyl)oxamic acid (0.095 g, 0.44 mmol)and (3S)-3-(leucinyl)amino-4-oxobutanoic acid (tert)-butyl estersemicarbazone (0.180 g, ca 0.41 mmol) in N-methylpyrrolidone(1.0mL)-CH₂Cl₂(1.0 mL) at 0° C. (ice balth) under nitrogen was addedhydroxybenzotriazole hydrate (0.100 g) followed by1-ethyl-3-(3′,3′-dimethyl-1′-aminopropyl)carbodiimide hydrochloride(0.100 g, 0.52 mmol). After stirring at 0° C. for 2 hrs and at roomtemperature for 16 hrs, the mixture was partitioned between EtOAc-water.The organic phase was washed with water, 5% KHSO₄, saturated NaHCO₃ andsaturated NaCl solutions, dried over anhydrous Na₂SO₄ and evaporated toa solid. The solid residue was triturated with Et₂O to give the titlecompound (0.231 g, 97%) as an off-white solid. TLC(MeOH—CH₂Cl₂; 5:95)Rf=0.32.

[0202] Part E:(3S)-3-[N-(N′-(1-Naphthyl)Oxamyl)Leucinyl]Amino-4-Oxobutanoic AcidSemicarbazone

[0203] To a suspension of(3S)-3-[N-(N′-(1-naphthyl)oxamyl)leucinyl]amino-4-oxobutanoic acid(tert)-butyl ester semicarbazone (0.212 g, 0.39 mmol) in CH₂Cl₂(2.0mL)-anisole(0.5 niL) at room temperature under nitrogen was addedtrifluoroacetic acid (2.0 mL). The resulting clear solution was stirredat room temperature for 3 hrs, evaporated to dryness and chased withtoluene-CH₂Cl₂ (1:1). The residue was triturated with Et₂O to give thetitle compound (0.181 g, 95%) as an off-white solid.TLC(AcOH—MeOH—CH₂Cl₂, 1:1:20) Rf=0.16.

[0204] Part F: (3S)-3-[N-(N′-(1-Naphthy)Oxamyl)Leucinyl]Amino-4-Oxobutanoic Acid

[0205] A suspension of (3S)-3-[N-(N′-(1-naphthyl)oxamyl)leucinyl]amino-4-oxobutanoic acidsemicarbazone (0.173 g, 0.36 mmol) in 37% aqueous formaldehyde(1.0mL)-acetic acid(I.0 mL)-methanol(3.0 mL) was stirred at room temperatureunder nitrogen for 18 hrs. The resulting clear solution was diluted withwater and the resulting white precipitate collected by suction andwashed with water. The combined aqueous filtrate was extracted withEtOAc. The extract was washed with water and saturated NaCl solution,dried over anhydrous Na₂SO₄ and evaporated to a glass. This was combinedwith the solid which was filtered from the aqueous mixture, taken up inCH₂Cl₂, filtered through Celite and evaporated to dryness. The crudeproduct was purified by dissolving the residue in CH₂Cl₂ andprecipitating with Et₂O-hexane. The precipitate was collected by suctionto give the title compound (0.129 g, 84%) as a white solid.TLC(AcOH—MeOH—CH₂Cl₂; 1:1:20) Rf=0.22. MS(ES) for C₂₂H₂₅N₃O₆ (MW427.46): positive 450(M+Na); negative 426(M−H)

EXAMPLE 2

[0206]

[0207] (3RS)-3-[N-(N′-(1-Naphthyl)Oxamyl)Leucinyl]Amino-5-Fluoro-4-Oxopentanoic Acid

[0208] Part A:(3RS,4RS)-3-[(N-Benzyloxycarbonyl)Leucinyl]Amino-5-Fluoro-4-HydroxypentanoicAcid, tert-Butyl Ester

[0209] To a solution of (3RS,4RS)-3-amino-5-fluoro-4-hydroxypentanoicacid, tert-butyl ester (0.230 g, 1.1 mmol, prepared as described inTetrahedron Letters 1994,35, 9693-9696) in CH₂Cl₂ (2.0 mL) at roomtemperature under nitrogen was added (N-benzyloxycarboyl)leucine,N-hydroxysuccinimde ester (0.402 g, 1.1 mmol). After stirring at roomtemperature for 16 hrs, the mixture was evaporated to dryness and theresidue purified by flash chromatography on silica gel eluting withEtOAc-hexane (1:2) to give the title compound (0.332 g, 66%) as acolorless, viscous oil. TLC(EtOAc-hexane; 2:1) Rf=0.51.

[0210] Part B: (3RS,4RS)-3-(Leucinyl)Amino-5-Fluoro-4-HydroxypentanoicAcid, tert-Butyl Ester, p-Toluenesulfonate Salt

[0211] To a solution of(3RS,4RS)-3-[(N-benzyloxycarbonyl)leucinyl]amino-5-fluoro-4-hydroxypentanoicacid, tert-butyl ester (0.332 g, 0.734 mmol) in MeOH (100 mL) was addedp-toluenesulsufonic acid hydrate (0.140 g, 0.737 mmol) and 10% Pd-C(0.033 g) and resulting mixture stirred under a hydrogen atmosphere(balloon) for 2 hrs. The mixture was filtered through Celite washing thefilter cake with CH₂Cl₂ and the combined filtrates evaporated todryness. The residue was chased with CH₂Cl₂ to give the title product(0.371 g) as a colorless foam.

[0212] Part C:(3RS,4RS)-3-[N-(N′-(1-Naphthyl)Oxamyl)Leucinyl]Amino-5-Fluoro-4-HydroxypentanoicAcid, tert-Butyl Ester

[0213] To a solution of N-(1-naphthyl)oxamic acid (0.161 g, 0.749 mmol,see Example 1, Part A) in N-methylpyrrolidone(1.5 mL)-CH₂Cl₂(1.5 mL) atroom temperature under nitrogen was addedO-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophate(0.313 g, 0.823 mmol). After stirring for 0.5 hrs, the mixture wastreated with a solution of(3RS,4RS)-3-(leucinyl)amino-5-fluoro-4-hydroxypentanoic acid, tert-butylester, p-toluenesulfonate salt (0.371 g, 0.749 mmol) anddiisopropylethylamine (0.39 mL, 2.25 mmol) in N-methylpyrrolidone(2.0mL)-CH₂Cl₂(2.0 mL). After stirring at room temperature for 16 hrs, themixture was partitioned between EtOAc-water. The organic phase waswashed with water, 5% KHSO₄, saturated NaHCO₃ and saturated NaClsolutions, dried over anhydrous Na₂SO₄ and evaporated to dryness. Theresidue was purified by flash chromatography on silica gel eluting withEtOAc-hexane (1:1) to give the title compound (0.213 g, 55%) as acolorless foam. TLC(Et₂O—CH₂Cl₂-hexane; 2:1:2, 2 developments) Rf=0.12.

[0214] Part D:(3RS)-3-[N-(N′-(1-Naphthyl)Oxamyl)Leucinyl]Amino-5-Fluoro-4-OxopentanoicAcid tert-Butyl Ester

[0215] To a solution of (3RS,4RS)-3-[N-(N′-(1-naphthyl)oxamyl)leucinyl]amino-5-fluoro-4-hydroxypentanoic acid, tert-butyl ester (0.163 g, 0.315mmol) in CH₂Cl₂ (3.0 mL) at room temperature was added Dess-Martinperiodinane (0.160 g, 0.378 mmol). After stirring at room temperaturefor 0.5 hrs, the mixture was diluted with EtOAc and washed with diluteNa₂S₂O₃, saturated NaHCO₃ and saturated NaCl solutions, dried overanhydrous Na₂SO₄ and evaporated to dryness. The residue was purified byflash chromatography on silica gel eluting with EtOAc-hexane (1:3) togive the title compound (0.155 g, 95%) as a white solid.TLC(Et₂O—CH₂Cl₂-hexane; 2:1:2, 2 developments) Rf=0.35. MS(ES) forC₂₇H₃₄FN₃O₆ (MW 515.57): positive 538(M+Na); negative 514(M−H).

[0216] Part E:(3RS)-3-[N-(N′-(1-Naphthyl)Oxamyl)Leucinyl]Amino-5-Fluoro-4-OxopentanoicAcid

[0217] To a solution of(3RS)-3-[N-(N′-(1-naphthyl)oxamyl)leucinyl]amino-5-fluoro-4-oxopentanoicAcid, tert-butyl ester (0.147 g, 0.285 mmol) in CH₂Cl₂(1.0mL)-anisole(0.5 mL) at room temperature under nitrogen was addedtrifluoroacetic acid (1.0 mL). The resulting clear solution was stirredat room temperature for 1 hr, evaporated to dryness and chased withtoluene-CH₂Cl₂ (1:1). The residue was triturated with Et₂O-hexane togive the title compound (0.100 g, 76%) as a white solid. MS(ES) forC₂₃H₂₆FN₃O₆ (MW 459.47): positive 482(M+Na); negative 458(M−H).

EXAMPLE 3

[0218]

[0219] (3RS)-3-[N-(N′-(1-Naphthyl)Oxamyl)Valinyl]Amino-5-Fluoro-4-Oxopentanoic Acid

[0220] Part A:(3RS,4RS)-3-[(N-Benzyloxycarbonyl)Valinyl]Amino-5-Fluoro-4-HydroxypentanoicAcid, tert-Butyl Ester

[0221] To a solution of (N-benzyloxycarbonyl)valine (0.332 g, 1.32 mmol)in CH₂Cl₂(7.0 mL) at 0° C. (ice bath) under nitrogen was addedhydroxybenzotriazole hydrate (0.219 g) followed by1-ethyl-3-(3′,3′-dimethyl-1′-aminopropyl)carbodiimide hydrochloride(0.317 g, 1.65 mmol). After stirring at 0° C. for 10 min, the mixturewas treated with (3RS,4RS)-3-amino-5-fluoro-4-hydroxypentanoic acid,tert-butyl ester (0.228 g, 1.1 mmol) and the reaction allowed to warm toroom temperature. After stirring at room temperature for 24 hrs, themixture was partitioned between EtOAc-water. The organic phase waswashed with water, 5% KHSO₄, saturated NaHCO₃ and saturated NaClsolutions, dried over anhydrous Na₂SO₄ and evaporated to dryness. Theresidue was purified by flash chromatography eluting with EtOAc-hexane(1:1) to give the title compound (0.423 g, 87%) as colorless glass.TLC(MeOH—CH₂Cl₂; 5:95) Rf=0.17.

[0222] Part B:(3RS)-3-[N-(N′-(1-Naphthyl)Oxamyl)Valinyl]Amino-5-Fluoro-4-OxopentanoicAcid

[0223] Starting with(3RS,4RS)-3-[(N-benzyloxycarbonyl)valinyl]amino-5-fluoro-4-hydroxypentanoicacid, tert-butyl ester and following the methods described in Example 2,Parts B through E gave the title compound as a white solid. MS(ES) forC₂₂H₂₄FN₃O₆ (MW 445.45): positive 468(M+Na), 484(M+K); negative444(M−H).

EXAMPLE 4

[0224]

[0225] (3S)-3-[N-(N′-(1-Naphthyl)Oxamyl)Valinyl]Amino-5(2′,6′-Dichlorobenzoyloxy)-4-Oxopentanoic Acid

[0226] Part A: [(N-Benzyloxycarbonyl)Valinyl]Aspartic Acid,β-tert-Butyl, α-Methyl Ester

[0227] To a solution of (N-benzyloxycarbonyl)valine (2.10 g, 8.36 mmol)in CH₂Cl₂(20 mL) at 0° C. (ice bath) under nitrogen was addedhydroxybenzotriazole hydrate (1.74 g) followed by1-ethyl-3-(3′,3′-dimethyl-1′-aminopropyl)carbodiimide hydrochloride(2.40 g, 12.5 mmol). After stirring at 0° C. for 10 min, the mixture wastreated with aspartic acid, β-tert-butyl, α-methyl ester hydrochloride(2.00 g, 8.34 mmol) and N-methylmorpholine (1.1 mL, 10 mmol), and thereaction allowed to warm to room temperature. After stirring at roomtemperature for 2.5 hrs, the mixture was concentrated and the residuepartitioned between EtOAc-water. The organic phase was washed withwater, 5% KHSO₄, saturated NaHCO₃ and saturated NaCl solutions, driedover anhydrous Na₂SO₄ and evaporated to give the title compound (3.55 g,97%) as a white solid after tituration with Et₂O-hexane.TLC(EtOAc-hexane; 1:1) Rf=0.48.

[0228] Part B: (YalinylAspartic Acid, β-tert-Butyl, α-Methyl Esterp-Toluenesulfonate Salt

[0229] To a solution of [(N-benzyloxycarbonyl)valinyl]aspartic acid,β-tert-butyl, α-methyl ester (3.55 g, 8.12 mmol) in MeOH (300 mL) wasadded p-toluenesulsufonic acid hydrate (1.55 g, 8.12 mmol) and 10% Pd-C(0.30 g) and resulting mixture stirred under a hydrogen atmosphere(balloon) for 2 hrs. The mixture was filtered through Celite washing thefilter cake with CH₂Cl₂ and the combined filtrates evaporated todryness. The residue was chased with CH₂Cl₂ to give the title product(3.85 g, quantitative) as a colorless foam.

[0230] Part C: [N-(N′-(1-Naphthyl)Oxamyl)Valinyl]Aspartic Acid,β-tert-Butyl, α-Methyl Ester

[0231] To a solution of N-(1-naphthyl)oxamic acid (0.683 g, 3.18 mmol,see Example 1, Part A) in N-methylpyrrolidone(7.0 mL)-CH₂Cl₂(7.0 mL) atroom temperature under nitrogen was addedO-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophate(1.329 g, 3.49 mmol). After stirring for 15 min, the mixture was treatedwith N-(valinyl)aspartic acid, 0-tert-butyl, x-methyl esterp-toluenesulfonate salt (1.506 g, 3.18 mmol) and diisopropylethylamine(1.66 mL, 9.53 mmol). After stirring at room temperature for 2 hrs, themixture was partitioned between EtOAc-water. The organic phase waswashed with water, 5% KHSO₄, saturated NaHCO₃ and saturated NaClsolutions, dried over anhydrous Na₂SO₄ and evaporated to dryness. Theresidue was purified by flash chromatography on silica gel eluting withEtOAc-hexane (1:1) to give the title compound (1. 1 53 g, 73%) as awhite solid. TLC(EtOAc-hexane; 2:1) Rf=0.48.

[0232] Part D: [N-(N′-(1-Naphthyl)Oxamyl)Valinyl]Aspartic Acid,β-tert-Butyl Ester

[0233] To a solution of [N-(N′-(1-naphthyl)oxamyl)valinyl] asparticacid, β-tert-butyl, α-methyl ester (0.490 g, 0.98 mmol) in dioxane (2.4mL) was added 1.0 N LiOH solution (1.0 mL, 1.0 mmol). After stirring atroom temperature for 1 hr, the mixture was acidified with 1.0 N HCl andextracted with EtOAc. The extract was washed with saturated NaClsolution, dried over anhydrous Na₂SO₄ and evaporated to give the titlecompound (0.481 g, quantitative) as a white solid. TLC(MeOH—CH₂Cl₂; 1:9)Rf=0.15.

[0234] Part E: (3S)-3-[N-(N′-(1-Naphthyl)Oxamyl)Valinyl]Amino-5-Diazo-4-Oxopentanoic Acidtert-Butyl Ester

[0235] To a solution of [N-(N′-(1-naphthyl)oxamyl)valinyl] asparticacid, β-tert-butyl ester (0.095 g, 0.20 mmol) and N-methylmorpholine (22μL, 0.20 mmol) in tetrahydrofuran (2.0 mL) at −10° C. (NaCl/ice bath)under nitrogen was added isobutyl chloroformate (28 μL, 0.22 mmol).After stirring at −10° C. for 0.5 hrs, the resulting mixed anhydride wastreated with excess diazomethane/Et₂O solution (prepared from 0.072 g,0.49 mmol of 1-methyl-3-nitro-1-nitrosoguanidine, 1.0 mL 40% KOH/1.0 mlEt₂O). After stirring at −10° C. for an additional 1 hr, the mixture wasconcentrated and the residue purified by flash chromatography on silicagel eluting with CH₂Cl₂—Et₂O-hexane (1:2:2) to give the title compound(0.062 g, 62%) as a white solid. TLC(EtOAc-hexane; 2:1) Rf=0.63.

[0236] Part F: (3S)-3-[N-(N′-(1-Naphthyl)Oxamyl)Valinyl]Amino-5-Bromo-4-Oxopentanoic Acidtert-Butyl Ester

[0237] To a solution of(3S)-3-[N-(N′-(1-naphthyl)oxamyl)valinyl]amino-5-diazo-4-oxopentanoicacid tert-butyl ester (0.135 g, 0.265 mmol) in tetrahydrofuran (3.0 mL)at 0° C. was added 48% aqueous HBr (30 μL, 0.27 mmol). Gas evolution wasobserved. After 15 min, the mixture was partitioned betweenEtOAc-saturated NaHCO₃, the organic phase washed with saturated NaClsolution, dried over anhydrous Na₂SO₄ and evaporated to give the titlecompound (0.147 g, quantitative) as a white solid. TLC(EtOAc-hexane;2:1) Rf=0.72.

[0238] Part G:(3S)-3-[N-(N′-(1-Naphthyl)Oxamyl)Valinyl]Amino-5-(2′,6′-Dichlorobenzoyloxy)-4-OxopentanoicAcid, tert-Butyl Ester

[0239] To a solution of (3S)-3-[N-(N′-(1-naphthyl)oxamyl)valinyl]amino-5-bromo-4-oxopentanoic acid tert-butyl ester (0.100 g,0.18 mmol) and 2,6-dichlorobenzoic acid (0.037 g, 0.20 mmol) indimethylformamide (1.0 mL) at room temperature under nitrogen was addedpotassium fluoride (0.031 g, 0.53 mmol). After stirring at roomtemperature for 16 hrs, the mixture was partitioned between EtOAc-water.The organic phase was washed with water, 5% KHSO₄, saturated NaHCO₃ andsaturated NaCl solutions, dried over anhydrous Na₂SO₄ and evaporated todryness. The residue was purified by flash chromatography on silica geleluting with EtOAc-hexane (1:1) to give the title compound (0.084 g,70%) as viscous oil. TLC(EtOAc-hexane; 2:1) Rf=0.71.

[0240] Part H:(S-3-[N-(N′-(1-Naphthyl)Oxamyl)Valinyl]AAmino-5-(2′,6′-Dichlorobenzoyloxy)-4-OxopentanoicAcid

[0241] To a solution of (3S)-3-[N-(N′-(1-naphthyl)oxamyl)valinyl]amino-5-(2′,6′-dichlorobenzoyloxy)-4-oxopentanoic acid,tert-butyl ester (0.084 g, 0.125 mmol) in CH₂Cl₂(1.0 mL)-anisole(0.5 mL)at room temperature under nitrogen was added trifluoroacetic acid (1.0mL). The resulting clear solution was stirred at room temperature for 1hr, evaporated to dryness and chased with toluene-CH₂Cl₂ (1:1). Theresidue was triturated with Et₂O to give the title compound (0.060 g,78%) as an off-white solid. MS(ES) for C₂₉H₂₇Cl₂N₃O₈ (MW 616.45):positive 638/640(M+Na); negative 614/616(M−H).

EXAMPLES 5-21

[0242] Starting with(3S)-3-[N-(N′-(1-naphthyl)oxamyl)valinyl]amino-5-bromo-4-oxopentanoicacid tert-butyl ester (see Example 4, Part F) and following the methodsdescribed in Example 4, Parts G through H, the compounds shown below inTable 3 were also prepared: TABLE 3

MS(ES) Ex. B Formula MW pos. neg. 5 CH₂O(2,6-diF-Ph) C₂₈H₂₇F₂N₃O₇ 555.53578(M + Na) 554(M − H) 6 CH₂O(2,4,6-triF-Ph) C₂₈H₂₆F₃N₃O₇ 573.52 596(M +Na) 572(M − H) 7 CH₂O(2,3,5,6-tetraF-Ph) C₂₈H₂₅F₄N₃O₇ 591.51 614(M + Na)590(M − H) 8 CH₂O(6-Me-2-pyron-4-yl) C₂₈H₂₉N₃O₉ 551.55 574(M + Na) 550(M− H) 9 CH₂O(2-Ph-5,6-benzopyran-4-on-3-yl) C₃₇H₃₃N₃O₉ 663.68 686(M + Na)662(M − H) 10 CH₂OPO(Me)Ph C₂₉H₃₂N₃O₈P 581.56 582(M + H) 580(M − H)604(M + Na) 694(M + TFA) 11 CH₂OPOPh₂ C₃₄H₃₄N₃O₈P 643.63 666(M + Na)642(M − H) 12 CH₂O(2-CF₃-pyrimidin-4-yl) C₂₇H₂₆F₃N₅O₇ 589.53 612(M + Na)588(M − H) 13 CH₂O(5-CO₂Me-isoxazol-3-yl) C₂₇H₂₈N₄O₁₀ 568.54 591(M + Na)567(M − H) 14 CH₂OPO(Me)(1-naphthyl) C₃₃H₃₄N₃O₈P 631.62 654(M + Na)630(M − H) 744(M + TFA) 15 CH₂O(4-hydroxy-Ph) C₂₈H₂₉N₃O₈ 535.55537/538(M + Na) 535/536/537 (M − H) 16 CH₂O(4-OCOOCHCHCH₃-Ph)C₃₂H₃₃N₃O₁₀ 619.63 637(M + NH₄) 618(M − H) 17 CH₂O(4-OCHCHCH₃-Ph)C₃₁H₃₃N₃O₈ 575.62 576(M + H) 574(M − H) 18 CH₂O(1-Naphthyl) C₃₂N₃₁N₃O₇569.6128 19 CH₂O(4-propyloxyphenyl) C₃₀H₃₅N₃O₆ 533.25 534(M + H) 551(M +NH₄) 20 CF₃ C₂₂H₂₂F₃N₃O₆ 481.15 482(M + H) 21 NH(CH₂)₂Ph C₂₉H₃₂N₄O₆532.23 533(M + H) 531(M − H)

EXAMPLE 22

[0243]

N-[1-(2-Benzyloxy-5-Oxo-Tetrahydro-Furan-3-Ylcarbamoyl)-Ethyl]-N′-(2-Tert-Butyl-Phenyl)-Oxalamide

[0244] Part A:3-(9H-Fluoren-9-ylmethoxycarbonylamino)-N-methoxy-N-methyl-suecinamicacid tert-butyl ester

[0245] To a solution of2-(9H-Fluoren-9-ylmethoxycarbonylamnino)-succinic acid 4-tert-butylester (5.00 g, 12.15 mmol) in CH₂Cl₂(50 mL) at 0° C. under N₂ was added1.2 equiv of 1-(3-Dimethylaminopropyl)-3-ethylcarbodiimide HCl (EDAC)(2.79 g, 14.58 mmol) and 1.1 equiv of HOBt.H₂O (2.05 g, 13.37 mmol). Themixture was stirred at 0° C. under N₂ for 15 min before addition of HClHN(OMe)Me (1.42 g, 14.58 mmol), followed by 4-Methylmorpholine (NM1M)(2.00 mL, 18.23 mmol). After stirring at 0° C. to room temperature for 5hrs, the mixture was partitioned between EtOAc/water. The organic phasewas washed with water, 5% KHSO₄, saturated NaHCO₃, and saturated NaClsolutions, dried over anhydrous Na₂SO₄ and evaporated to give crudetitle compound. The residue was purified by flask column chromatographyon silica gel eluting with EtOAc/Hexane (20%) to give the title compound(4.62 g, 85%) as a white foam. TLC (40%) Et(Ac/Hexane) R_(f)=0.44; ¹HNMR(CDCl₃) δ 7.75 (d, J=7.5 Hz, 2H), 7.50 (t, J=6.9 Hz, 2H), 7.40 (t, J=7.5Hz, 2H), 7.33-7.28 (dt, J=7.5, 1.2 Hz, 2H), 5.71 (d, J=9.0 Hz, 1H), 5.04(bq, J=7.8 Hz, 1H), 4.36 (d, J=7.2 Hz, 2H), 4.23 (t J=7.2 Hz, 1H), 3.79(s, 3H), 3.24 (s, 3H), 2.78-2.71 (dd, J=15.0, 5.4 Hz, 1H1), 2.62-254(15.0, 6.9 Hz, 1H), 1.45 (s, 9H). MS(ES) for C₂₃H₂₅NO₆ (MW=454.52):positive 455 (MH⁺).

[0246] Part B: 3-(9H-Fluoren-9-ylmethoxycarbonylamino)-4-oxo-butyricacid tert-butyl ester

[0247] To a suspension of3-(9H-Fluoren-9-ylmethoxycarbonylamino)-N-methoxy-N-methyl-succinamicacid tert-butyl ester (4.57 g, 10.04 mmol) in Ethyl ether (30 mL) at −5°C. was added dropwise 0.5 equiv of 1.0M Lithium aluminum hydride (LAH)in ethyl ether (5.02 mL, 5.02 mmol). After stirring at −5° C. to roomtemperature for 1 hr, the reaction mixture was treated with 5% KHSO₄(200 mL), stirred for 5 min and then partitioned between EtOAc/water.The organic phase was washed with saturated NaCl solution (200 mL),dried over Na₂SO₄ and evaporated to dryness. The residue was purified byflask column chromatography on silica gel eluting with EtOAc/Hexane(20%) to give the title compound (2.18 g, 55%/o) as a clear oil. TLC(30% EtOAc/Hexane) R_(f)=0.35; ¹HNMR (CDCl₃) δ 9.65 (s, 1H), 7.77 (d,J=7.2 Hz, 2H), 7.50 (d, J=7.5 Hz, 21), 7.41 (t, J=7.5 Hz, 2H), 7.33 (dt,J=7.5, 1.2 Hz, 2H), 5.88 (d, J=7.2 Hz, 11H), 4.50-4.36 (m, 2H), 4.24 (t,J 6.6 Hz, 1H), 3.54-3.45 (m, 1H), 3.00-2.93 (dd, J=17.4, 4.8 Hz, 11H),2.82-274 (dd, J=17.7, 5.1s Hz, 1H), 1.45 (s, 9H). MS(ES) for C₂₃H₂₅NO₅(MW=395.45): positive 396 (MH⁺).

[0248] Part C:4,4-Bis-benzyloxy-3-(9H-fluoren-9-ylmethoxycarbonylamino)-butyric acidtert-butyl ester

[0249] To a suspension of3-(9H-Fluoren-9-ylmethoxycarbonylamino)-4-oxo-butyric acid tert-butylester (1.24 g, 3.14 mmol) in Dichloromethane (10 ML) at room temperaturewas added 0.20 equiv TsOH.H₂O (0.12 g, 0.63 mmol) and followed bydropwise 5.0 equiv of Benzyl alcohol (1.62 mL, 15.68 mmol). Afterstirring room temperature for 18 hrs, the reaction mixture was filteredthrough celite and the filtrate was concentrated in vacuo to obtain acrude title compound. The residue was purified by flask columnchromatography on silica gel eluting with EtOAc/Hexane (10-40%) to givethe title compound (0.75 g, 40%) as clear oil. TLC (40% EtOAc/Hexane)R_(f)=0.68; ¹HNMR (CDCl₃) δ 7.76 (d, J=7.2 Hz, 2H), 7.59 (d, J=7.5 Hz,2H), 7.39 (t, J=7.5 Hz, 2H), 7.35-7.27 (m, 12H), 5.37 (d, J=9.6 Hz,1IH), 4.73-4.52 (m, 5H), 4.35 (d, J=6.6 Hz, 2H), 4.24-4.16 (q, J=6.6 Hz,2H), 2.55-2.50 (m, 2H), 1.39 (s, 9H). MS(ES) for C₃₇H₃₉NO₉ (MW=593.71):positive 611 (M+NH₄ ⁺).

[0250] Part D:4,4-Bis-benzyloxy-3-{2-[(2-tert-butyl-phenylaminooxalyl)-aminol-propionylamino]-butvricacid tert-butyl ester

[0251] 4,4-Bix-benzyloxy-3-(9H-fluoren-9-ylmethoxycarbonylamino)-butyricacid tert-butyl ester (0.63 g, 1.06 mmol) was stirredin 5%Piperidine/DMlF (3 mL) at room temperature. After stirring at roomtemperature for 15 min, the reaction mixture was treated with Water (20mL), stirred for 5 min and then partitioned between EtOAc/water. Theorganic phase was washed with Water (3×20 mL), saturated NaCl solution(3×20 mL), dried over Na₂SO₄ and evaporated to dryness to afford crude3-Amino-4,4-bis-benzyloxy-butyric acid tertbutyl ester (1). The cruderesidue 1 was ready to be used for coupling reaction as a crude product.

[0252] To a solution of(2S)-2-[(2-tert-butyl-phenylaminooxalyl)-amino]-propanoic acid (0.37 g,1.27 mmol) in CH₂Cl₂/NMP (1:1, 3 mL) at room temperature under N2 wasadded 1.5 equiv HBTU (0.61 g, 1.59 mmol). The mixture solution wasstirred at room temperature under N₂ for 45 min before addition of 1,followed by Et₃ N (0.44 mL, 3.18 mmol). After stirring at roomtemperature for 18 hrs, the mixture was partitioned between EtOAc-water.The organic phase was washed with water, 5% KHSO₄, saturated NaHCO₃, andsaturated NaCl solutions, dried over anhydrous Na₂SO₄ and evaporated togive crude title compound. The residue was purified by flask columnchromatography on silica gel eluding with EtOAc/Hexane (20%) to give thetitle compound (0.29 g, 42%) as a yellow foam. TLC (30% EtOAc/Hexane)R_(f)=0.59; ¹HNMR (CDCl₃) δ 9.53 (s, 1H), 8.06 (dd, J=8.1, 3.6 Hz, 1H),8.01 (dt, J=8.1, 1.2 Hz, 1H), 7.42 (d, J=7.5 Hz, 1H), 7.37-7.25 (m,11H), 7.17 (tt, J=7.5, 1.2 Hz, 1H), 6.62-6.53 (dd, J=16.2, 9.0 Hz, 1-),4.72-4.35 (m, 7H), 2.66 (d, J=6.0 Hz, 2H), 1.45 (s, 9H), 1.38 (d, J=4.2Hz, 3H); MS(ES) for C₃₇H₄₇N₃O₇ (MW-645.79): negative 654 ([M−H]⁻).

[0253] Part E:N-[1-(2-Benzyloxy-5-oxo-tetrahydro-furan-3-ylcarbamoyl)-ethyl]-N′-(2-tert-butyl-phenyl)-oxalamide

[0254]4,4-Bis-benzyloxy-3-{2-[(2-tert-butyl-phenylaminooxalyl)-amino]-propionylamino}-butyricacid tert-butyl ester (0.16 g, 0.25 mmol) was stirred in 205% TFA/CH₂Cl₂(2 mL) at room temperature. After stirring at room temperature for 15min., the reaction mixture was concentrated to dryness to afford crudetittle compounds. The residue was purified by flask columnchromatography on silica gel eluding with EtOAc/Hexane (20-40%) to givethe title compounds (0.09 g, 74%) as white foam.

[0255] Major diastereomer: TLC (40% EtOAc/Hexane) R_(f)=0.29; ¹HNMR(CDCl₃) δ 9.53 (d, J=7.2 Hz, 1H), 8.09 (t, J=7.8 Hz, 1H), 7.93 (m, 1H),7.42 (d, J=9.0 Hz, 1H), 7.35-7.15 (m, 7H), 6.70 (d, J=8.1 Hz, 1H), 5.45(d, J=5.1 Hz, 1H), 4.90-4.41 (m, 4H), 3.06-2.83 (m, 1H), 2.53-2.35 (m,1H), 1.47 (d, J=7.2 Hz, 3H), 1.44 (s, 9H); MS(ES) for C₂₆H₃₁N₃O₆(MW=481.54): positive 482 (MH+).

[0256] Minor diastereomer: TLC (40% EtOAc/Hexane) R_(f)=0.22; ¹HNMR(CDCl₃) δ 9.52 (s, 1H), 8.02-7.93 (m, 2H), 7.44 (dd, J=8.1, 1.5 Hz, 1H),7.36-8.28 (m, 6H), 7.19 (dt,J=9.30, 1.5Hz, IH))6.67 (d,J=7.8 Hz, 11H),5.54 (d, J=5.4 Hz, 1H), 4.92-4.44 (m, 4H), 2.95-2.85 (m, 1H), 2.55-2.44(m, 1H), 1.49 (d, J=7.2 Hz, 3H), 1.44 (s, 9H); MS(ES) for C₂₆H₃₁N₃O₆(MW=481.54): positive 482 (MH+).

EXAMPLE 23

[0257]

[0258](3S)-3-[N-(N′-(1-Naphthyl)Oxamyl)Leucinyl]Amino-5-(Diphenylphosphinyloxy)-4-OxopentanoicAcid

[0259] Part A: [(N-Benzyloxycarbonyl)Leucinyl]Aspartic Acid β-tert-Butylα-Methyl Ester

[0260] To a solution of (N-benzyloxycarbonyl)leucine,N-hydroxysuccinimide ester (4.54 g, 12.5 mmol) and aspartic acid,β-tert-butyl, α-methyl ester hydrochloride (3.00 g, 12.5 mmol) in CH₂Cl₂(20 mL) at room temperature under nitrogen was added N-methylmorpholine(1.65 mL, 15 mmol). After stirring at room temperature for 18 hrs, themixture was partitioned between EtOAc-water. The organic phase waswashed with 5% KHSO₄, saturated NaHCO₃ and saturated NaCl solutions,dried over anhydrous Na₂SO₄ and evaporated to give the title compound(5.56 g, 99%) as viscous oil. TLC(EtOAc-hexane; 1:1) Rf=0.48.

[0261] Part B:(3S)-3-[N-(N′-(1-Naphthyl)Oxamyl)Leucinyl]Amino-5-Bromo-4-OxopentanoicAcid tert-Butyl Ester

[0262] Starting with [(N-benzyloxycarbonyl)leucinyl]aspartic acid,β-tert-butyl, α-methyl ester and following the methods described inExample 4, Parts B through F, gave the title compound as a white solid.TLC(CH₂Cl₂-Et₂O-hexane; 1:2:2) Rf=0.32.

[0263] Part C:(3S)-3-[N-(N′-(1-Naphthyl)Oxamyl)Leucinyl]Amino-5-(Diphenylphopshinyloxy)-4-OxopentanoicAcid, tert-Butyl Ester

[0264] To a solution of (3 S)-3-[N-(N′-(1-naphthyl)oxamyl)leucinyl]amino-5-bromo-4-oxopentanoic acid tert-butyl ester (0.108 g,0.187 mmol) and diphenylphosphinic acid (0.046 g, 0.21 mmol) indimethylformamide (1.0 mL) at room temperature under nitrogen was addedpotassium fluoride (0.033 g, 0.58 mmol). After stirring at roomtemperature for 48 hrs, the mixture was partitioned between EtOAc-water.The organic phase was washed with water, 5% KHSO₄, saturated NaHCO₃ andsaturated NaCl solutions, dried over anhydrous Na₂SO₄ and evaporated todryness. The residue was purified by flash chromatography on silica geleluting with CH₂Cl₂—Et₂O-hexane (1:2:2) to give the title compound(0.114 g, 85%) as a white solid. TLC(EtOAc-hexane; 2:1) Rf=0.26.

[0265] Part D:(3S)-3-[N-(N′-(1-Naphthyl)Oxamyl)Leucinyl]Amino-5-(Diphenylphosphinyloxy)-4-OxopentanoicAcid

[0266] To a solution of (3S)-3-[N-(N′-(1-naphthyl)oxamyl)leucinyl]amino-5-(diphenylphosphinyloxy)-4-oxopentanoic acid, tert-butylester (0.114 g, 0.16 mmol) in CH₂Cl₂(1.0 mL)-anisole(0.5 mL) at roomtemperature under nitrogen was added trifluoroacetic acid (1.0 mL). Theresulting clear solution was stirred at room temperature for 1 hr,evaporated to dryness and chased with toluene-CH₂Cl₂ (1:1). The residuewas triturated with Et₂O-hexane to give the title compound (0.062 g,59%) as an off-white solid. MS(ES) for C₃₄H₃₄N₃O₈P (MW 657.66): positive680(M+Na); negative 656(M−H).

EXAMPLES 24-27

[0267] Starting with(3S)-3-[N-(N′-(1-naphthyl)oxamyl)leucinyl]amino-5-bromo-4-oxopentanoicacid tert-butyl ester (see Example 23, Part B) and following the methodsdescribed in Example 23, Parts C through D, the compounds shown below inTable 4 were also prepared: TABLE 4

MS(ES) Ex. B Formula MW pos. neg. 24 CH₂OCO(2,6-diCl-Ph) C₃₀H₂₉Cl₂N₃O₈630.48 652/654(M + Na) 628/630(M − H) 25 CH₂O(2,4,6-triF-Ph)C₂₉H₂₈F₃N₃O₇ 587.55 610(M + Na) 586(M − H) 26 CH₂O(2,3,5,6-tetraF-Ph)C₂₉H₂₇F₄N₃O₇ 605.54 628(M + Na) 604(M − H) 27 CH₂OPO(Me)Ph C₃₀H₃₄N₃O₈P595.59 596(M + H) 594(M − H) 618(M + Na) 708(M + TFA)

EXAMPLES 28-77

[0268] Following the general methods described in Example 4, Parts Athrough H substituting (N-benzyloxycarbonyl)alanine for(N-benzyloxycarbonyl)valine in Part A, the appropriate oxamic acid forN-(1-naphthyl)oxamic acid in Part C, and the appropriate acid or phenolfor 2,6-dichlorobenzoic acid in Part G, the compounds shown below inTable 5 were also prepared: TABLE 5

MS(ES) Ex. R¹ B Formula MW Pos. neg. 28 (2-Ph)Ph CH₂O(2-F-Ph)C₂₈H₂₆FN₃O₇ 535.53 558(M + Na) 534(M − H) 29 (2-Ph)PhCH₂OCO(2,6-di-Cl-Ph) C₂₉H₂₅Cl₂N₃O₈ 614.44 652/654(M + K) 612/614(M − H)30 (2-Ph)Ph CH₂OPOPh₂ C₃₄H₃₂N₃O₈P 641.61 664(M + Na) 640(M − H) 680(M +K) 31 (2-t-Bu)Ph CH₂O(2-F-Ph) C₂₆H₃₀FN₃O₇ 515.54 516(M + H) 514(M − H)538(M + Na) 554(M + K) 32 (2-t-Bu)Ph CH₂OPOPh₂ C₃₂H₃₆N₃O₈P 621.63644(M + Na) 620(M − H) 666(M + K) 33 1-naphthyl-CH₂CH₂O(2,3,5,6-tetra-F-Ph) C₂₇H₂₃F₄N₃O₇ 577.48 600(M + Na) 576(M − H)616(M + K) 34 1-naphthyl-CH₂ CH₂OCO(2,6-di-Cl-Ph) C₂₈H₂₅Cl₂N₃O₈ 602.42624/626(M + Na) 600/602(M − H) 640/642(M + K) 714/716 (M + TFA) 351-naphthyl-CH₂ CH₂OPOPh₂ C₃₃H₃₂N₃O₈P 629.60 652(M + Na) 628(M − H)668(M + K) 36 (2-tBu)Ph CH₂O(2,3,5,6-tetra-F-4-Br)Ph) C₂₆H₂₆BrF₄N₃O₇648.40 646(M − H) 37 (2-tBu)Ph CH₂OCO(2,6-di-Cl-Ph) C₂₇H₂₉Cl₂N₃O₈ 594.45616/618(M + Na) 492/594(M − H) 38 (2-1)Ph CH₂OCO(2,6-di-Cl-Ph)C₂₃H₂₀Cl₂IN₃O₈ 664.24 39 (2-1)Ph CH₂OCO(2,6-di-Cl-Ph) C₂₃H₂₀Cl₂FN₃O₈556.33 40 (2,5-di-tBu)Ph CH₂O(2,3,5,6-tetra-F-Ph) C₃₀H₃₅F₄N₃O₇ 625.62626(M + H) 624(M − H) 648(M + Na) 41 (2-Cl)Ph CH₂OPOPh₂ C₂₈H₂₇CIN₃O₈P599.96 598(M − H) 42 (2-Cl)Ph CH₂OCO(2,6-di-Cl-Ph) C₂₃H₂₀Cl₃N₃O₈ 572.79572/574/576 570/572/574 (M + H) (M − H) 43 (4-F)Ph CH₃OPOPh₂C₂₈H₂₇FN₃O₈P 583.51 606(M + H) 582(M − H) 44 (4-F)PhCH₂OCO(2,6-di-Cl-Ph) C₂₃H₂₀Cl₂FN₃O₈ 556.33 578/480(M + Na) 554/556(M −H) 45 (2-morpholin-4-yl-5-trifluoromethyl)Ph CH₂OPOPh₂ C₃₅H₃₅F₆N₄O₁₁P832.65 741(M + Na) 717(M − H) 46 (2-pyrrolidin-1-yl-5-trifluoromethyl)PhCH₂OPOPh₂ C₃₅H₃₅F₆N₄O₁₀P 816.65 725(M + Na) 701(M − H) 47 (2-tBu)PhCH₂OPOPh₂ C₃₂H₃₆N₃O₈P 621.63 644(M + Na) 620(M − H) 485,6,7.8-tetrahydro-1-naphthyl CH₂OPOPh₂ C₃₂H₃₄N₃O₈P 619.61 620(M + H)618(M − H) 642(M + Na) 49 5,6,7.8-tetrahydro-1-naphthylCH₂OCO(2,6-di-Cl-Ph) C₂₇H₂₇Cl₂N₃O₈ 592.43 614/616(M + Na) 590/592(M − H)50 5,6,7.8-tetrahydro-1-naphthyl CH₂O(2,3,5,6-tetra-F-ph) C₂₆H₂₅F₄N₃O₇567.49 590(M + Na) 566(M − H) 51 PhCH₂ CH₂O(2,3,5,6-tetra-F-Ph)C₂₃H₂₃Cl₂N₃O₈ 552.37 550/552(M − H) 52 PhCH₂ CH₂OCO(2,6-di-Cl-Ph)C₂₄H₂₃Cl₂N₃O₈ 552.37 550/552(M − H) 53 1-adamantanyl CH₂OPOPh₂C₃₂H₃₈N₃O₈P 623.64 646(M + Na) 622(M − H) 54 1-adamantanylCH₂OCO(2,6-di-Cl-Ph) C₂₇H₃₁Cl₂N₃O₈ 596.46 594/596(M − H) 55 cyclo-hexylCH₂OCO(2,6-di-Cl-Ph) C₂₃H₂₇Cl₂N₃O₈ 544.39 566(M + Na) 56 cyclo-hexylCH₂OPOPh₂ C₂₈H₃₄N₃O₈P 571.57 594(M + Na) 570(M − H) 610(M + K) 57(2-Cl)Ph CH₂OPOPh₂ C₂₈H₂₇ClN₃O₈P 5999.96 622/624(M + Na) 598/600(M − H)58 (2-Cl)Ph CH₂OCO(2,6-di-Cl-Ph) C₂₃H₂₀Cl₃N₃O₈ 572.79 594(M + Na) 59(2,5-di-6Bu)Ph CH₂OPOPh₂ C₃₆H₄₄N₃O₈P 677.73 700(M + Na) 676(M − H) 60(2,5-di-6Bu)Ph CH₂OCO(2,6-di-Cl-Ph) C₃₁H₃₇Cl₂N₃O₈ 650.55 650/652(M + H)648/650(M − H) 672/674(M + Na) 61 1,2,3,4-tetrahydro-1-naphthylCH₂OPOPh₂ C₃₂H₃₄N₃O₈P 619.61 620(M + H) 618(M − H) 642(M + Na) 621,2,3,4-tetrahydro-1-naphthyl CH₂OCO(2,6-di-Cl-Ph) C₂₇H₂₇Cl₂N₃O₈ 592.43614/616(M + Na) 590/592(M − H) 63 1-adamantanyl CH₂O(2,3,5,6-tetra-F-Ph)C₃₀H₂₃F₄N₃O₇ 613.52 636(M + Na) 612(M − H) 64 1-adamantanyl CH₂OPOPh₂C₃₆H₃₂N₃O₈ 665.64 688(M + Na) 664(M − H) 65 1-adamantanylCH₂OCO(2,6-di-Cl-Ph) C₃₁H₂₅Cl₂N₃O₈ 638.46 660/662(M + Na) 636/638(M − H)66 (2-F)Ph CH₂O(2,3,5,6-tetra-F-Ph) C₂₂H₁₈F₅N₃O₇ 531.39 554(M + Na)530(M − H) 67 (2-F)Ph CH₂OPOPh₂ C₂₈H₂₇FN₃O₈P 583.51 606(M + Na) 582(M −H) 68 (2-F)Ph CH₂OCO(2,6-di-Cl-Ph) C₂₃H₂₀Cl₂FN₃O₈ 556.33 578/580(M + Na)554/556(M − H) 69 (4-n-heptyl)Ph CH₂OPOPh₂ C₃₅H₄₂N₃O₈P 663.71 686(M +Na) 662(M − H) 70 (4-n-heptyl)Ph CH₂OCO(2,6-di-Cl-Ph) C₃₀H₃₅Cl₂N₃O₈636.53 658/660(M + Na) 634/636(M − H) 71 Ph(CH₂)₂CH₂O(2,3,5,6-tetra-F-Ph) C₂₄H₂₃F₄N₃O₇ 541.46 542(M + H) 540(M − H)564(M + Na) 72 Ph(CH₂)₂ CH₂OPOPh₂ C₃₀H₃₂N₃O₈P 593.57 594(M + H) 592(M −H) 616(M + Na) 566/568(M + H) 73 Ph(CH₂)₂ CH₂OCO(2,6-di-Cl-Ph)C₂₅H₂₅Cl₂N₃O₈ 566.39 566/568(M + H) 564/566(M − H) 588/590(M + Na)714(M + Na) 74 (2-1)Ph CH₂OPOPh₂ C₂₈H₂₇IN₃O₈P 691.41 714(M + Na) 690(M −H) 75 (2-1)Ph CH₂OCO(2,6-di-Cl-Ph) C₂₃H₂₀Cl₂IN₃O₈ 664.24 686/688(M + Na)662/664(M − H) 776(M + TFA) 76 (2-tBu)Ph CH₂OCO(2,6-di-Cl-Ph)C₂₇H₂₉Cl₂N₃O₈ 594.45 616/618(M + Na) 592/594(M − H) 77 (2-PhCH₂)PhCH₂O(2,3,5,6-tetra-F-Ph) C₂₉H₃₀N₃O₈P 579.57 578(M − H)

EXAMPLE 78

[0269]

[0270](3S)-3-[N-(N′-(1-Naphthylmethyl)Oxamyl)Valinyl]Amino-5-(2,′,3′,5′,6′-Tetrafluorophenoxy)-4-OxopentanoicAcid

[0271] Part A: [(N-Benzyloxycarbonyl)Valinyl]Aspartic Acid, β-tert-ButylEster

[0272] To a suspension of aspartic acid P-tert-butyl ester (3.784 g, 20mmol) in acetonitrile (200 mL) at room temperture under nitrogen wasadded bis(trimethylsilyl)acetamide (9.9 niL, 40 mmol). After stirring atroom temperature for 30 min, the resulting clear solution was treatedwith (N-benzyloxycarbonyl)valine N-hydroxysuccinimide ester (6.97 g, 20mmol). After stirring at room temperature for an additional 18 hrs, themixture was treated with water (20 mL), concentrated on a rotovap andthen partitioned between EtOAc/water. The organic phase was washed withwater, 5% KHSO₄ and saturated NaCl solutions, dried over anhydrousNa₂SO₄ and evaporated to a dryness. Trituration with Et₂O-hexane gavethe title compound (8.37 g, 99%) as a white solid. TLC(EtOAc-hexane;1:1) Rf=0.06.

[0273] Part B:(3S)-3-[(N-Benzyloxycarbonyl)Valinyl]Amino-5-Bromo-4-Oxopentanoic Acidtert-Butyl Ester

[0274] A solution of [(N-benzyloxycarbonyl)valinyl]aspartic acid,β-tert-butyl ester (8.37 g, 19.9 mmol) and N-methylmorpholine (3.50 mL,32 mmol) in tetrahydrofuran (100 mL) at −10° C. (NaCl/ice bath) undernitrogen was treated dropwise with isobutyl chloroformate (3.87 mL, 29.8mmol). After stirring at −10° C. for 20 min, the mixture was filtered(sinctered glass) into a pre-cooled receiver (ice bath) washing thefilter cake with additional tetrahydrofuran (approx.30 mL). The combinedfiltrate was treated with excess diazomethane/Et₂O solution (preparedfrom 7.32 g, 50 mmol of 1-methyl-3-nitro-1-nitrosoguanidine, 40 mL 40%KOH/65 ml Et₂O) at 0° C. (ice bath) under nitrogen. After stirring at 0°C. for 15 min and at room temperature for 30 min, the reaction mixturewas again cooled to 0° C. and treated with 48% HBr(10 mL, 60mmol)/acetic acid(10 mL). After stirring at 0° C. for 15 min and at roomtemperature for 30 min, the mixture was partitioned between EtOAc-water.The organic phase was washed with water, saturated NaHCO₃, and saturatedNaCl solutions dried over anhydrous Na₂SO₄ and evaporated to a dryness.Trituration with hexane gave the crude title compound (9.71 g, 98%) as awhite solid. TLC(EtOAc-hexane; 1:1) Rf=0.63.

[0275] Part C:(3S)-3-[(N-Benzyloxycarbonyl)Valinyl]Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-OxopentanoicAcid tert-Butyl Ester

[0276] To a solution of(3S)-3-[(N-benzyloxycarbonyl)valinyl]amino-5-bromo-4-oxopentanoic acidtert-butyl ester (9.71 g, 19.4 mmol) and 2,3,5,6-tetrafluorophenol (3.65g, 22 mmol) in tetrahydrofuran (20 niL) at room temperature undernitrogen was added potassium fluoride (2.91 g, 50 mmol). After stirringat room temperature for 4 hrs, the mixture was diluted with EtOAc(approx.100 mL), washed with saturated NaHCO₃ and saturated NaClsolutions, dried over anhydrous Na₂SO₄ and evaporated to a dryness. Theresidue was purified by flash chromatography on silica gel eluting withEtOAc-hexane (1:3) to give the title compound (9.19 g, 79%) as a whitesolid after trituration with Et₂O-hexane. TLC(EtOAc-hexane; 1:1)Rf=0.70.

[0277] Part D:(3S,4RS)-3-[(N-Benzyloxycarbonyl)Valinyl]Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-HydroxypentanoicAcid tert-Butyl Ester

[0278] To a solution of(3S)-3-[(N-benzyloxycarbonyl)valinyl]amino-5-(2′,3′,5′,6′-tetrafluorophenoxy)-4-oxopentanoicacid tert-butyl ester (9.19 g, 15.7 mmol) in MeOH(200mL)/tetrahydrofuran(200 mL) at 0° C. under nitrogen was added sodiumborohydride (0.594 g, 15.7 mmol). After stirring at 0° C. for 1 hr, themixture was concentrated and the residue partitioned between EtOAc-halfsaturated NH₄Cl solution. The organic phase was washed with saturatedNaHCO₃ and saturated NaCl solutions, dried over anhydrous Na₂SO₄ andevaporated to a dryness. The residue was purified by flashchromatography on silica gel eluting with EtOAc-hexane (1:3) to give thetitle compound (7.99 g, 87%) as a white solid. TLC(EtOAc-hexane; 1:1)Rf=0.54.

[0279] Part E:(3S,4RS)-3-(alinyl)Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-HydroxypentanoicAcid tert-Butyl Ester

[0280] To a solution of(3S,4RS)-3-[(N-benzyloxycarbonyl)valinyl]amino-5-(2′,3′,5′,6′-tetrafluorophenoxy)-4-hydroxypentanoicacid tert-butyl ester (7.99 g, 13.6 mmol) in MeOH (130 mL) was added 10%Pd-C (0.80 g) and resulting mixture stirred under a hydrogen atmosphere(balloon) for 2 hrs. The mixture was filtered through Celite washing thefilter cake with CH₂Cl₂ and the combined filtrates evaporated todryness. The residue purified by flash chromatography on silica geleluting with EtOAc-hexane (1:3) then methanol to give the title compound(5.13 g, 83%) as a viscous oil. TLC(EtOAc-hexane; 1:1) Rf=0.07.

[0281] Part F: (3S,4RS)-3-[N-(N′-(1-Naphthylmethyl)Oxamyl)Valinyl]Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-Hydroxypentanoic Acidtert-Butyl Ester

[0282] To a solution of N-(1-naphthylmethyl)oxamic acid (0.051 g, 0.22mmol, prepared from 1-naphthylmethylamine by the method described inExample 1, Part A) in N-methylpyrrolidone(1.0 mL)-CH₂Cl₂(1.0 mL) at roomtemperature under nitrogen was addedO-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophate(0.092 g, 0.24 mmol). After stirring for 15 min, the mixture was treatedwith(3S,4RS)-3-(valinyl)amino-5-(2′,3′,5′,6′-tetrafluorophenoxy)-4-hydroxypentanoicacid tert-butyl ester (0.100 g, 0.22 mmol) and diisopropylethylamine(115 μL, 0.66 mmol). After stirring at room temperature for 16 hrs, themixture was partitioned between EtOAc-water. The organic phase waswashed with water, 5% KHSO₄, saturated NaHCO₃ and saturated NaClsolutions, dried over anhydrous Na₂SO₄ and evaporated to give the crudetitle compound (0.157 g, 100%) as a viscous oil. TLC(EtOAc-hexane; 1:1)Rf=0.44.

[0283] Part G:(3S)-3-[N-(N′-(1-Naphthylmethyl)Oxamyl)Valinyl]Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-OxopentanoicAcid tert-Butyl Ester

[0284] To a solution of(3S,4RS)-3-[N-(N′-(1-naphthylmethyl)oxamyl)valinyl]-amino-5-(2′,3′,5′,6′-tetrafluorophenoxy)-4-hydroxypentanoicacid tert-butyl ester (0.157 g, ca 0.22 mmol) in dimethylsulfoxide (5mL) at room temperature under nitrogen was added Dess-Martin Periodinane(0.600 g, 1.42 mmol). After stirring at room temperature for 16 hrs, themixture was partitioned between EtOAc-water. The organic phase waswashed with saturated NaHCO₃ and saturated NaCl solutions, dried overanhydrous Na₂SO₄ and evaporated to a dryness. The residue (0.175 g) waspurified by flash chromatography on silica gel eluting with EtOAc-hexane(3:7) to give the title compound (0.111 g, 77%) as a white solid.TLC(EtOAc-hexane; 1:1) Rf=0.58.

[0285] Part H:(3S)-3-[N-(N′-(1-Naphthylmethyl)Oxamyl)Valinyl]Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-OxopentanoicAcid

[0286] To a solution of(3S)-3-[N-(N′-(1-naphthylmethyl)oxamyl)valinyl]amino-5-(2′,3′,5′,6′-tetrafluorophenoxy)-4-oxopentanoicacid, tert-butyl ester (0.108 g, 0.16 mmol) in CH₂Cl₂(2.0mL)-anisole(0.1 mL)-water(0.05 mL) at room temperature under nitrogenwas added trifluoroacetic acid (2.0 mL). The resulting clear solutionwas stirred at room temperature for 2 hr, evaporated to dryness andchased with toluene-CH₂Cl₂ (1:1). The residue was triturated with Et₂Oto give the title compound (0.098 g, 100%) as a white solid. MS(ES) forC₂₉H₂₇F₄N₃O₇ (MW 605.54): positive 628(M+Na); negative 604(M−H).

EXAMPLES 79-125

[0287] Starting with(3S,4RS)-3-(valinyl)amino-5-(2′,3′,5′,6′-tetrafluorophenoxy)-4-hydroxypentanoicacid tert-butyl ester (see Example 78, Part E) and following the methodsdescribed in Example 78, Parts F through H, the compounds shown below inTable 6 were also prepared: TABLE 6

MS(ES) Ex. R¹ Formula MW pos. neg. 79 PhCH₂ C₂₅H₂₅F₄N₃O₇ 555.48 556(M +H) 554(M − H) 578(M + Na) 80 Ph(CH₂)₂ C₂₆H₂₇F₄N₃O₇ 569.51 592(M + Na)568(M − H) 81 Ph₂CH C₃₁H₂₉F₄N₃O₇ 631.58 654(M + Na) 630(M − H) 82 PhC₂₄H₂₃F₄N₃O₇ 541.46 564(M + Na) 540(M − H) 83 (2-Ph)Ph C₃₀H₂₇F₄N₃O₇617.55 640(M + Na) 616(M − H) 730(M + TFA) 84 (2-PhCH₂)Ph C₃₁H₂₉F₄N₃O₇631.58 654(M + Na) 630(M − H) 85 (3-PhO)Ph C₃₀H₂₇F₄N₃O₈ 633.55 634(M +H) 632(M − H) 656(M + Na) 86 4-Cl-1-naphthyl C₂₈H₂₄ClF₄N₃O₇ 625.96648/650(M + Na) 624/626(M − H) 87 2-anthryl C₃₂H₂₇F₄N₃O₇ 641.57 642(M +H) 640(M − H) 88 2-benzimidazolyl C₂₅H₂₃F₄N₅O₇ 581.48 582(M + H) 580(M −H) 604(M + Na) 89 1-adamantanyl C₂₈H₃₃F₄N₃O₇ 599.58 600(M + H) 598(M −H) 90 (2-F)Ph C₂₄H₂₂F₅N₃O₇ 559.45 582(M + Na) 558(M − H) 672(M + TFA) 91(4-F)Ph C₂₄H₂₂F₅N₃O₇ 559.45 582(M + Na) 558(M − H) 672(M + TFA) 92(2-CF₃)Ph C₂₅H₂₂F₇N₃O₇ 609.45 632(M + Na) 608(M − H) 722(M + TFA) 93(2-t-Bu)Ph C₂₈H₃₁F₄N₃O₇ 597.56 620(M + Na) 596(M − H) 710(M + TFA) 94(4-n-heptyl)Ph C₃₁H₃₇F₄N₃O₇ 639.64 662(M + Na) 638(M − H) 95 (2-CH₃O)PhC₂₅H₂₅F₄N₃O₈ 571.48 594(M + Na) 570(M − H) 96 (2-PhO)Ph C₃₀H₂₇F₄N₃O₈633.55 656(M + Na) 632(M − H) 746(M + TFA) 97 2-naphthyl C₂₈H₂₅F₄N₃O₇591.51 614(M + Na) 590(M − H) 98 5,6,7,8-tetrahydro-1-naphthylC₂₈H₂₉F₄N₃O₇ 595.55 618(M + Na) 594(M − H) 99 1-anthryl C₃₂H₂₇F₄N₃O₇641.57 664(M + Na) 640(M − H) 100 2-pyridinyl C₂₃H₂₂F₄N₄O₇ 542.44543(M + H) 541(M − H) 101 4-pyridinyl C₂₃H₂₂F₄N₄O₇ 542.44 543(M + H)541(M − H) 102 2,3,5,6-tetrafluoro-4-pyridinyl C₂₃H₁₈F₈N₄O₇ 614.40615(M + H) 613(M − H) 103 2-pyrazinyl C₂₂H₂₁F₄N₅O₇ 543.43 544(M + H)542(M − H) 104 1,2,3,4-tetrahydro-1-naphthyl C₂₈H₂₉F₄N₃O₇ 595.55 596(M +H) 594(M − H) 618(M + Na) 708(M + TFA) 634(M + K) 105 (2-Cl)PhC₂₄H₂₂ClF₄N₃O₇ 575.90 598/600(M + Na) 574/576(M − H) 106 (2-Br)PhC₂₄H₂₂BrF₄N₃O₇ 620.35 644/642(M + Na) 620/618(M − H) 734/732(M + TFA)107 (2-I)Ph C₂₄H₂₂F₄IN₃O₇ 667.35 690(M + Ma) 666(M − H) 706(M + K)780(M + TFA) 108 (2,6-di-F)Ph C₂₄H₂₂F₆N₃O₇ 577.44 600(M + Na) 576(M − H)690(M + TFA) 109 (2,5-di-t-Bu)Ph C₃₂H₃₉F₄N₃O₇ 653.67 654(M + H) 652(M −H) 676(M + Na) 688(M + Cl) 692(M + K) 766(M + TFA) 110 5-indanylC₂₇H₂₇F₄N₃O₇ 581.52 604(M + Na) 580(M − H) 620(M + K) 694(M + TFA) 111(3,4,5-tri-MeO)PhCH₂ C₂₈H₃₁F₄N₃O₁₀ 645.56 646(M + H) 644(M − H) 668(M +Na) 684(M + K) 112 methyl C₁₉H₂₁F₄N₃O₇ 479.38 502(M + Na) 478(M − H)592(M + TFA) 113 n-heptyl C₂₅H₃₃F₄N₃O₇ 563.55 586(M + Na) 562(M − H)602(M + K) 676(M + TFA) 114 t-octyl C₂₆H₃₅F₄N₃O₇ 577.57 600(M + Na)576(M − H) 115 cyclo-hexyl C₂₄H₂₉F₄N₃O₇ 547.50 548(M + H) 546(M − H)570(M + Na) 660(M + TFA) 586(M + K) 116 5-Ph-3-pyrazolyl C₂₇H₂₅F₄N₅O₇607.52 630(M + Na) 606(M − H) 646(M + K) 117 (2-F-4-I)Ph C₂₄H₂₁F₅IN₃O₇685.34 686(M + H) 684(M − H) 708(M + Na) 720(M + Cl) 724(M + K) 118(2,3,4,5-tetra-F)Ph C₂₄H₁₉F₈N₃O₇ 613.41 614(M + H) 612(M − H) 636(M +Na) 726(M + TFA) 652(M + K) 119 (2,3,4,6-tetra-F)Ph C₂₄H₁₉F₈N₃O₇ 613.41614(M + H) 612(M − H) 636(M + Na) 726(M + TFA) 652(M + K) 120(2,3,5,6-tetra-Cl)Ph C₂₄H₁₉Cl₄F₄N₃O₇ 679.23 700/702/704(M + Na)676/678/680(M − H) 716/718/720(M + K) 790/792/794 (M + TFA) 121(2,3,4,5,6-penta-F)Ph C₂₄H₁₈F₉N₃O₇ 631.40 654(M + Na) 630(M − H) 670(M +K) 666(M + Cl) 122 Ph₂N C₃₀H₂₈F₄N₄O₇ 632.57 633(M + H) 631(M − H)655(M + Na) 745(M + TFA) 123 PHCH₂(Ph)N C₃₁H₃₀F₄N₄O₇ 646.59 647(M + H)645(M − H) 669(M + Na) 681(M + Cl) 685(M + K) 124 PhCH₂O C₂₅H₂₅F₄N₃O₇571.48 594(M + Na) 570(M − H) 684(M + TFA) 125 5-quinolinyl C₂₉H₂₅F₇N₄O₉706.53 593(M + H) 591(M − H)

EXAMPLE 126

[0288]

[0289](3S)-3-[N-(N′-(2-Tert-Butylphenyl)Oxamyl)Alaninyl]Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4)-OxopentanoicAcid

[0290] Part A: [(N-Benzyloxycarbonyl)Alaninyl]Aspartic Acid,β-tert-Butyl Ester

[0291] To a suspension of aspartic acid β-tert-butyl ester (3.784 g, 20mmol) in dimethylformamide (150 mL) at room temperture under nitrogenwas added bis(trimethylsilyl)-trifluoroacetamide (10.6 niL, 40 mmol).After stirring at room temperature for 30 min, the resulting clearsolution was treated with (N-benzyloxycarbonyl)alanineN-hydroxysuccinimide ester (6.406 g, 20 mmol). After stirring at roomtemperature for an additional 48 hrs, the mixture was treated with water(20 mL), stirred for 15 min and then partitioned between EtOAc/water.The organic phase was washed with water, 5% KHSO₄ and saturated NaClsolutions, dried over anhydrous Na₂SO₄ and evaporated to a dryness. Theresidue was dissolved in Et₂O and extracted with saturated NaHCO₃. Theaqueous extract was acidified (pH 2.0) with concentrated HCl andextracted with EtOAc. The EtOAc extract was washed with saturated NaClsolution, dried over anhydrous Na₂SO₄ and evaporated to a give the titlecompound (6.463 g, 82%) as a white foam. TLC(EtOAc-hexane-AcOH; 70:30:2)Rf=0.50.

[0292] Part B:(3S,4RS)-3-(Alaninyl)Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-HydroxypentanoicAcid tert-Butyl Ester

[0293] Starting with [(N-benzyloxycarbonyl)alaninyl]aspartic acid,β-tert-butyl ester and following the methods described in Example 28,Parts B through E gave the title compound as a colorless, viscous oil.TLC(EtOAc-hexane; 1:1) Rf=0.06.

[0294] Part C: (3S,4RS)-3-[N-(N′-(2-tert-Butylphenyl)Oxamy1)Alaninyl]Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-HydroxypentanoicAcid tert-Butyl Ester

[0295] To a solution of N-(2-tert-butylphenyl)oxamic acid (0.041 g, 0.19mmol, prepared from 2-tert-butylaniline by the method described inExample 1, Part A) in CH₂Cl₂(6.0 mL) at 0° C. under nitrogen was addedhydroxybenzotriazole hydrate (0.030 g) followed by1-ethyl-3-(3′,3′-dimethyl-1′-aminopropyl)- carbodiinmide hydrochloride(0.050 g, 0.26 mmol). After stirring at 0° C. for 10 min, the mixturewas treated with(3S,4RS)-3-(alaninyl)amino-5-(2′,3′,5′,6′-tetrafluorophenoxy)-4-hydroxypentanoicacid tert-butyl ester (0.079 g, 0.19 mmol) and N-methylmorpholine (22μL, 0.20 mmol). After stirring at room temperature for 16 hrs, themixture was partitioned between EtOAc-water. The organic phase waswashed with water, 5% KHSO₄, saturated NaHCO₃ and saturated NaClsolutions, dried over anhydrous Na₂SO₄ and evaporated to give the crudetitle compound (0.090 g, 77%) as a viscous oil. TLC(EtOAc-hexane; 1:1)Rf=0.70.

[0296] Part D:(3S)-3-[N-(N′-(2-tert-Butylphenyl)Oxamy)Alaninyl]Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-OxopentanoicAcid tert-Butyl Ester

[0297] To a solution of(3S,4RS)-3-[N-(N′-(2-tert-butylphenyl)oxamyl)alaninyl]amino-5-(2′,3′,5′,6′-tetrafluorophenoxy)-4-hydroxypentanoic acidtert-butyl ester (0.0.092 g, ca 0.15 mmol) in CH₂Cl₂ (6.5 mL) at roomtemperature under nitrogen was added iodobenzene diacetate (0.188 g,0.58 mmol) followed by a catalytic amount of2,2,6,6-tetramethyl-1-piperidinyloxy free radical (TEMPO, 0.0046 g, 0.03mmol). After stirring at room temperature for 16 hrs, the mixture waspartitioned between EtOAc-water. The organic phase was washed withsaturated NaHCO₃ and saturated NaCl solutions, dried over anhydrousNa₂SO₄ and evaporated to a dryness. The residue (0.096 g) was purifiedby preparative layer chromatography on silica gel eluting withEtOAc-hexane (3:7) to give the title compound (0.071 g, 77%) as acolorless glass. TLC(EtOAc-hexane; 2:3) Rf=0.60.

[0298] Part E:(3S)-3-[N-(N′-(2-tert-Butylphenyl)Oxamyl)Alaninyl]Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-OxopentanoicAcid

[0299] To a solution of(3S)-3-[N-(N′-(2-tert-butylphenyl)oxamyl)alaninyl]amino-5-(2′,3′,5′,6′-tetrafluorophenoxy)-4-oxopentanoicacid, tert-butyl ester (0.071 g, 0.11 mmol) in CH₂Cl₂(2.5mL)-anisole(0.05 mL) at room temperature under nitrogen was addedtrifluoroacetic acid (1.5 mL). The resulting clear solution was stirredat room temperature for 1 hr, evaporated to dryness and chased withtoluene-CH₂Cl₂ (1:1). The residue (0.061 g) was purified by preparativelayer chromatography on silica gel eluting with MeOH—CH₂Cl₂ (1:9) togive the title compound (0.044 g, 69%) as a colorless glass. MS(ES) forC₂₆H₂₇F₄N₃O₇ (MW 569.51): positive 570(M+H); negative 568(M−H).

EXAMPLES 127-178

[0300] Starting with(3S,4RS)-3-(alaninyl)amino-5-(2′,3′,5′,6′-tetrafluorophenoxy)-4-hydroxypentanoicacid tert-butyl ester (see Example 79, Part B) and following the methodsdescribed in Example 79, Parts C through E, the compounds shown below inTable 7 were also prepared: TABLE 7

MS(ES) Ex. R¹ Formula MW pos. neg. 127 (2-CF₃)Ph C₂₃H₁₈F₇N₃O₇ 581.40604(M + Na) 580(M − H) 128 (2-Ph)Ph C₂₈H₂₃F₄N₃O₇ 589.50 612(M + Na)588(M − H) 129 (2-PhCH₂)Ph C₂₉H₂₅F₄N₃O₇ 603.53 604(M + H) 602(M − H) 130(2-PhO)Ph C₂₈H₂₃F₄N₃O₈ 605.50 628(M + Na) 604(M − H) 131 (3-PhO)PhC₂₈H₂₃F₄N₃O₈ 605.50 628(M + Na) 604(M − H) 1325,6,7,8-tetrahydro-1-naphthyl C₂₆H₂₅F₄N₃O₇ 567.49 590(M + Na) 566(M − H)133 1-naphthyl C₂₆H₂₁F₄N₃O₇ 563.46 586(M + Na) 562(M − H) 608(M + K) 134Ph C₂₂H₁₉F₄N₃O₇ 513.40 552(M + K) 512(M − H) 135 (2,6-di-F)PhC₂₂H₁₇F₆N₃O₇ 549.38 572(M + Na) 548(M − H) 662(M + TFA) 136 (4-Ph)PhC₂₈H₂₃F₄N₃O₇ 589.50 — 588(M − H) 137 (4-MeO)Ph C₂₃H₂₁F₄N₃O₈ 543.43582(M + K) 542(M − H) 138 Ph₂CH C₂₉H₂₅F₄N₃O₇ 603.53 642(M + K) 602(M −H) 139 4-pyridinyl C₂₂H₂₀F₄N₄O₉ 560.42 515(M + H) 513(M + H) 1402-pyridinyl C₂₁H₁₈F₄N₄O₇ 514.39 515(M + H) 141 (2-Cl)Ph C₂₂H₁₈ClF₄N₃O₇547.85 142 (2,3,4,5-tetra-Cl)Ph C₂₂H₁₅F₈N₃O₇ 585.36 584(M − H) 1435-indanyl C₂₅H₂₃F₄N₃O₇ 553.47 552(M − H) 144 (2-Br)Ph C₂₂H₁₈BrF₄N₃O₇592.30 590/592(M − H) 145 (2,3,5,6-tetra-F)Ph C₂₂H₁₅Cl₄F₄N₃O₇ 651.18648/650/652 (M − H) 146 1-anthryl C₃₀H₂₃F₄N₄O₇ 613.52 636(M + Na) 612(M− H) 147 PhCH₂ C₂₃H₂₁F₄N₃O₇ 527.43 148 Ph(CH₂)₂ C₂₄H₂₃F₄N₃O₇ 541.46542(M + H) 540(M − H) 564(M + Na) 149 (2-F)Ph C₂₂H₁₈F₅N₃O₇ 531.39554(M + Na) 530(M − H) 150 (4-F)Ph C₂₂H₁₈F₅N₃O₇ 531.39 554(M + Na) 530(M− H) 151 (2-pyrrolidin-1-yl)Ph C₂₈H₂₇F₇N₄O₉ 696.53 583(M + H) 581(M − H)695(M + TFA) 152 (2-morpholin-4-yl)Ph C₂₈H₂₇F₇N₄O₁₀ 712.53 599(M + H)597(M − H) 711(M + TFA) 153 (2-morpholin-4-yl-5-trifluoromethyl)PhC₂₉H₂₆F₁₀N₄O₁₀ 780.53 689(M + Na) 664(M − H) 154(2-pyrrolidin-1-yl-5-trifluoromethyl)Ph C₂₉H₂₆F₁₀N₄O₉ 764.53 651(M + H)649(M − H) 673(M + Na) 155 PhCH₂ C₂₃H₂₁F₄N₃O₇ 527.43 594(M + Na) 590(M −H) 156 Adamantan-1-yl C₂₆H₂₉F₄N₃O₇ 571.52 594(M + Na) 570(M − H) 157cyclo-hexyl C₂₂H₂₅F₄N₃O₇ 519.45 558(M + K) 518(M − H) 158 (2-Cl)PhC₂₂H₁₈ICF₄N₃O₇ 547.85 570/572 546/548(M − H) (M + Na) 159(2,5,di-t-Bu)Ph C₃₀H₃₅F₄N₃O₇ 625.61 1601,2,3,4-tetrahydro-naphthalen-1-yl C₂₆H₂₅F₄N₃O₇ 567.49 590(M + Na) 566(M− H) 161 (4-n-heptyl)Ph C₂₉H₃₃F₄N₃O₇ 611.49 634(M + Na) 610(M − H) 162(2-1)Ph C₂₂H₁₈F₄IN₃O₇ 639.30 662(M + Na) 638(M − H) 163naphthalen-1yl-methyl C₂₇H₂₃F₄N₃O₇ 577.49 600(M + Na) 576(M − H) 164pyrrolidin-1-yl C₂₀H₂₂F₄N₄O₇ 506.14 507.22 (M + H) 165 1-pyrrolidineC₂₀H₂₁F₄N₃O₇ 491.13 509(M + NH₄) 492(M + H) 166 1-piperidineC₂₁H₂₃F₄N₃O₇ 505.15 504(M + H) 167 piperidin-1-yl C₂₃H₂₈F₄N₄O₉ 580.18519(M − H) 168 (2-tBu-5-AcNH)Ph C₂₈H₃₀F₄N₄O₈ 626.2 625(M − H) 169(2-tBu-5-NH(CO(CH₂)3NHGmoc)Ph C₄₅H₄₅F₄N₅O₁₀ 891.31 890(M − H) 170(2,3,5,6-tetra-F)Ph C₂₁H₁₆F₈N₄O₆ 572.09 573(M + H) 171 (2-(3-CH₃O-Ph))PhC₂₉H₂₅F₄N₃O₈ 619.16 642(M + Na) 618(M − H) 172 (2-(4-CH₃O-Ph))PhC₂₉H₂₅F₄N₃O₈ 619.16 620(M + H) 618(M − H) 173 (2-(2-CH₃O-Ph))PhC₂₉H₂₅F₄N₃O₈ 619.16 620(M + H) 618(M − H) 174 (2-(1-naphthyl))PhC₃₂H₂₅F₄N₃O₇ 639.55 662(M + Na) 638(M − H) 175 [2-{(3-CH₃)Ph}]PhC₂₉H₂₅F₄N₃O₇ 603.19 626(M + Na) 602(M − H) 176 [2-{(4-CH₃)Ph}]PhC₂₉H₂₅F₄N₃O₇ 603.19 626(M + Na) 602(M − H) 177 [2-{(2-CH₃)Ph}]PhC₂₉H₂₅F₄N₃O₇ 603.19 626(M + Na) 602(M − H)

EXAMPLE 178

[0301]

[0302] (3S)-3-[N-(N′-(2′-Phenoxyphenyl)Oxamyl)Cyclohexylalaninyl]Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-Oxopentanoic Acid

[0303] Part A: (3S)-3-(N-Benzyloxycarbonyl)Amino-5-Bromo-4-OxopentanoicAcid tert-Butyl Ester

[0304] A solution of (N-benzyloxycarbonyl)aspartic acid, P-tert-butylester (2.28 g, 7.06 mmol) and N-methylmorpholine (0.85 m,L 7.7 mmol) intetrahydrofuran (40 mL) at −10° C. (NaCl/ice bath) under nitrogen wastreated dropwise via syringe with isobutyl chloroformate (1.1 mL, 8.5mmol). After stirring at −10° C. for 20 min, the mixture was filtered(sinctered glass) into a pre-cooled receiver (ice bath) washing thefilter cake with additional tetrahydrofuran (approx.10 mL). The combinedfiltrate was treated with excess diazomethane/Et₂O solution (preparedfrom 3.10 g, 21 mmol of 1-methyl-3-nitro-1-nitrosoguanidine, 20 mL 40%KOH/10 ml Et₂O) at 0° C. (ice bath) under nitrogen. After stirring at 0°C. for 15 min and at room temperature for 30 min, the reaction mixturewas again cooled to 0° C. and treated with 48% HBr(2.0 mL, 12mmol)/acetic acid(2.0 mL). After stirring at 0° C. for 15 min and atroom temperature for 15 min, the mixture was partitioned betweenEtOAc-water. The organic phase was washed with water, saturated NaHCO₃,and saturated NaCl solutions dried over anhydrous Na₂SO₄ and evaporatedto a dryness. Trituration with hexane gave the crude title compound(3.32 g) as a yellow oil. TLC(EtOAc-hexane; 1:1) Rf=0.60 (intermediatediazoketone Rf=0.52).

[0305] Part B: (3S,4RS)-3-(N-Benzyloxycarbonyl)Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-HydroxypentanoicAcid tert-Butyl Ester

[0306] To a solution of(3S)-3-(N-benzyloxycarbonyl)amino-5-bromo-4-oxopentanoic acid tert-butylester (0.857 g, 2.14 mmol) and 2,3,5,6-tetrafluorophenol (0.410 g, 2.45mmol) in dimethylformamide (5.0 mL) at room temperature under nitrogenwas added potassium fluoride (0.40 g, 6.9 mmol). After stirring at roomtemperature for 16 hrs, the mixture was diluted with EtOAc, washed withsaturated NaHCO₃ and saturated NaCl solutions, dried over anhydrousNa₂SO₄ and evaporated to a to give the crude tetrafluorophenoxymethylketone (1.08 g, 98%) as a yellow, viscous oil. TLC(EtOAc-hexane;1:1)Rf=0.57.

[0307] To a solution of the above crude ketone (1.08 g, ca 2.14 mmol) inethanol (10 mL) at 0° C. under nitrogen was added sodium borohydride(0.057 g, 1.5 mmol). After stirring at 0° C. for 1 hr. the excessreducing agent was discharged by treatment with acetone (1.0 mL), themixture concentrated and the residue partitioned between EtOAc-halfsaturated NH₄Cl solution. The organic phase was washed with saturatedNaHCO₃ and saturated NaCl solutions, dried over anhydrous Na₂SO₄ andevaporated to a dryness. The residue was purified by flashchromatography on silica gel eluting with EtOAc-hexane (1:3) to give thetitle compound (1.012 g, 94%) as a colorless oil. TLC(EtOAc-hexane; 1:1)Rf=0.48.

[0308] Part C:(3S,4RS)-3-[(N-9-Fluorenylmethoxycarbonyl)Cyclohexylalaninyl]Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-HydroxypentanoicAcid tert-Butyl Ester

[0309] To a solution of(3S,4RS)-3-(N-benzyloxycarbonyl)amino-5-(2′,3′,5′,6′-tetrafluorophenoxy)-4-hydroxypentanoicacid tert-butyl ester (1.012 g, 2.08 mmol) in MeOH (25 mL) was added 10%Pd-C (0.30 g) and resulting mixture stirred under a hydrogen atmosphere(balloon) for 4 hrs. The mixture was filtered through Celite washing thefilter cake with CH₂Cl₂ and the combined filtrates evaporated to givethe crude amine (0.682 g, 93%) as aviscous oil. TLC(MeOH—CH₂Cl₂; 5:95)Rf=0.21.

[0310] To a solution of (N-9-fluorenylmethoxycarbonyl) cyclohexylalanine(0.763 g, 1.94 mmol) in CH₂Cl₂(10 mL) at 0° C. (ice bath) under nitrogenwas added hydroxybenzotriazole hydrate (0.282 g) followed by1-ethyl-3-(3′,3′-dimethyl-1′-aminopropyl)carbodiimide hydrochloride(0.447 g, 2.33 mmol). After stirring at 0° C. for 10 min, the mixturewas treated with the above crude amine (0.682 g, ca 1.93 mmol) and thereaction allowed to warm to room temperature. After stirring at roomtemperature for 3 hrs, the mixture was partitioned between EtOAc-water.The organic phase was washed with water, 5% KHSO₄, saturated NaHCO₃ andsaturated NaCl solutions, dried over anhydrous Na₂SO₄ and evaporated todryness. The residue was purified by flash chromatography eluting withEtOAc-hexane (1:2) to give the title compound (1.028 g, 73%) as yellowfoam. TLC(EtOAc-hexane; 1:2) Rf=0.20.

[0311] Part D:(3S,4RS)-3-[Cyclohexyalaninyl]Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-HydroxypentanoicAcid tert-Butyl Ester

[0312] A mixture of(3S,4RS)-3-[(N-9-fluorenylmethoxycarbonyl)cyclohexylalaninyl]amino-5-(2′,3′,5′,6′-tetrafluorophenoxy)-4-hydroxypentanoic acidtert-butyl ester (1.028 g, 1.4 mmol) and 10%piperidine/dimethylformamide (3.0 mL) was stirred at room temperatureunder nitrogen for 2 hrs. The mixture was diluted with CH₂Cl₂, washedwith water and saturated NaHCO₃ solution, dried over anhydrous Na₂SO₄and evaporated to dryness. The residue was purified by flashchromatography eluting with isopropanol-CH₂Cl₂ (7:93) to give the titlecompound (0.561 g, 78%) as a white solid. TLC(MeOH—CH₂Cl₂; 5:95)Rf=0.43.

[0313] Part E:(3S.4RS)-3-[N-(N′-(2′-Phenoxyphenyl)Oxamy1)Cyclohexylalaninyl]-Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-HydroxypentanoicAcid tert-Butyl Ester

[0314] To a solution of N-(2-phenoxyphenyl)oxamic acid (0.064 g, 0.25mmol, prepared from 2-phenoxyaniline by the method described in Example1, Part A) and(3S,4RS)-3-[cyclohexylalaninyl]amino-5-(2′,3′,5′,6′-tetrafluorophenoxy)-4-hydroxypentanoicAcid tert-butyl ester (0.124 g, 0.245 mmol) in CH₂Cl₂(5.0 mL) at 0° C.(ice bath) under nitrogen was added hydroxybenzotriazole hydrate (0.051g) followed by 1-ethyl-3-(3′,3′-dimethyl-1′-aminopropyl)carbodiimidehydrochloride (0.061 g, 0.32 mmol). After stirring at 0° C. for 10 minand at room temperature for 18 hrs, the mixture was partitioned betweenEtOAc-water. The organic phase was washed with water, 5% KHSO₄,saturated NaHCO₃ and saturated NaCl solutions, dried over anhydrousNa₂SO₄ and evaporated to give the crude title compound (0.194 g) asyellow foam. TLC(EtOAc-hexane; 1:2) Rf=0.40.

[0315] Part F:(3S)-3-[N-(N′-(2′-Phenoxypheny1)Oxamyl)Cyclohexylalaniny1]-Amino-S-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-OxoxypentanoicAcid tert-Butyl Ester

[0316] To a solution of crude(3S,4RS)-3-[N-(N′-(2′-phenoxyphenyl)oxamyl)cyclohexylalaninyl]amino-5-(2′,3′,5′,6′-tetrafluorophenoxy)-4-hydroxypentanoicacid tert-butyl ester (0.194 g, ca 0.245 mmol) in CH₂Cl₂ (5 mL) at roomtemperature under nitrogen was added Dess-Martin Periodinane (0.150 g,0.35 mmol). After stirring at room temperature for 2 hrs, the mixturewas diluted with EtOAc, washed with 1.0 M Na₂S₂O₃, saturated NaHCO₃ andsaturated NaCl solutions, dried over anhydrous Na₂SO₄ and evaporated toa dryness. The residue was purified by flash chromatography on silicagel eluting with EtOAc-hexane (1:3) to give the title compound (0.142 g,80%) as a colorless, viscous oil. TLC(EtOAc-hexane; 1:2) Rf=0.50.

[0317] Part G:(3S)-3-[N-(N′-(2′-Phenoxyphenyl)Oxamyl)Cyclohexylalaninyl]-Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-OxoxypentanoicAcid

[0318] To a solution of(3S)-3-[N-(N′-(2′-phenoxyphenyl)oxamyl)cyclohexylalaninyl]amino-5-(2′,3′,5′,6′-tetrafluorophenoxy)-4-oxopentanoic acid, tert-butylester (0.142 g, 0.19 mmol) in CH₂Cl₂(2.0 mL) at room temperature undernitrogen was added trifluoroacetic acid (1.0 mL). The resulting clearsolution was stirred at room temperature for 0.5 hr, evaporated todryness and chased with toluene-CH₂Cl₂ (1:1) to give the title compound(0.123 g, 93%) as a white foam. MS(ES) for C₃₄H₃₃F₄N₃O₈ (MW 687.64):positive 688(M+H), 710(M+Na), 726(M+K); negative 686(M−H), 800(M+TFA).

EXAMPLES 179-181

[0319] Starting with(3S,4RS)-3-[cyclohexylalaninyl]amino-5-(2′,3′,5′,6′-tetrafluoro-phenoxy)-4-hydroxypentanoicacid tert-butyl ester (see Example 178, Part D) and following themethods described in Example 178, Parts E through G the compounds shownbelow in Table 8 were also prepared: TABLE 8

MS(ES) Ex. R1 Formula MW pos. neg. 179 (2-Ph)Ph C₃₄H₃₃F₄N₃O₇ 671.64 672670(M − H) (M + H) 784 694 (M + TFA) (M + Na) 180 (2-PhCH₂)PhC₃₅H₃₅F₄N₃O₇ 685.67 708 684(M − H) (M + Na) 798 (M + TFA) 181 1-naphthylC₃₂H₃₁F₄N₃O₇ 645.61 668 644(M − H) (M + Na) 758 (M + TFA)

EXAMPLE 182

[0320]

[0321](3S)-3-[N-(N′-(5,6,7,8-Tetrahydro-1-Naphthyl)Oxamyl)-Cyclohexylalaninyl]Amino-5-(2′,6′-Dichlorobenzoyloxy)-4-Oxopentanoic Acid

[0322] Part A: Aspartic Acid, β-tert-Butyl α-Methyl Esterp-Toluenesulfonate Salt

[0323] To a solution of N-(benzyloxycarbonyl)-L-aspartic acid,β-tert-butyl ester (10.57 g, 32.7 mmol) in methanol(20 mL)-CH₂Cl₂(30 mL)at 0° C. (ice bath) was added portionwise a 2.0 M solution of(trimethylsilyl)diazometbane in hexanes (20 mL, 40 mmol). After stirringat 0° C. for 45 min, the excess reagent was quenced with formic acid(1.0 rL). The mixture was washed with saturated NaHCO₃ solution, driedover anhydrous Na₂SO₄ and evaporated to a pale yellow oil (11.34 g).

[0324] The crude product (11.34 g, ca 32.7 mmol) was taken up inmethanol (100 mL), treated with p-toluenesulfonic acid mono hydrate(6.20 g, 32.6 mmol) and 10% Pd-C (0.5 g) and stirred under a hydrogenatmosphere (balloon) for 3 hrs. The mixture was filtered through Celiteand concentrated to give the title compound as a white solid (12.68 g).

[0325] Part B: [(N-Benzyloxycarbonyl)Cyclohexylalaninyl]Aspartic Acid,β-tert-Butyl α-Methyl Ester

[0326] To a solution of (N-benzyloxycarbonyl)-cyclohexylalaninedicyclohexylamine salt (0.866 g, 1.77 mmol) in CH₂Cl₂(10 mL) at 0° C.(ice bath) under nitrogen was added hydroxybenzotriazole hydrate (0.100g) followed by 1-etbyl-3-(3′,3′-dimethyl-1′-aminopropyl)carbodiimidehydrochloride (0.41 g, 2.14 mmol). After stirring at 0° C. for 10 min,the mixture was treated with aspartic acid, β-tert-butyl, α-methyl esterp-toluenesulfonate salt (0.665 g, 1.77 mmol) and N-methylmorpholine (0.2mL, 1.8 mmol), and the reaction allowed to warm to room temperature.After stirring at room temperature for 2.5 hrs, the mixture wasconcentrated and the residue partitioned between EtOAc-water. Theorganic phase was washed with water, 5% KHSO₄, saturated NaHCO₃ andsaturated NaCl solutions, dried over anhydrous Na₂SO₄ and evaporated toan oil. Purification by flash chromatography on silica gel eluting withEtOAc-hexane (1:3) gave the title compound (0.764 g, 88%) as a viscousoil. TLC(EtOAc-hexane; 1:2) Rf=0.46.

[0327] Part C:(3S)-3-[N-(N′-(5,6,7,8-Tetrahydro-1-Naphthyl)Oxamyl)-Cyclohexylalaninyl]Amino-5-(2′,6′-Dichlorobenzoyloxy)-4-OxopentanoicAcid

[0328] Starting with [(N-benzyloxycarbonyl)cyclohexyl-alaninyl]asparticacid, β-tert-butyl, α-methyl ester and following the general methodsdescribed in Example 4, Parts B through H, gave the title compound as awhite solid. MS(ES) for C₃₃H₃₇Cl₂N₃O₈ (MW 674.58): positive696/698(M+Na); negative 672/674(M−H), 786/788(M+TFA).

EXAMPLES 183-189

[0329] Starting with [(N-benzyloxycarbonyl)cyclohexyl-alaninyl]asparticacid, β-tert-butyl, α-methyl ester (see Example 182, Part B), andfollowing the general methods described in Example 4, Parts B through H,the compounds shown below in Table 9 were also prepared: TABLE 9

MS(ES) Ex. R1 B Formula MW pos. neg. 183 5,6,7,8-tetrahydro-1-naphthylCH₂O(2,3,5,6-tetra-F-Ph) C₃₂H₃₅F₄N₃O₇ 649.64 672(M + Na) 648(M − H) 1845,6,7,8-tetrahydro-1-naphthyl CH₂OPO(Me)Ph C₃₃H₄₂N₃O₈P 639.68 662(M +Na) 638(M − H) 752(M + TFA) 185 5,6,7,8-tetrahydro-1-naphthyl CH₂OPOPh₂C₃₈H₄₄N₃O₈P 701.75 724(M + Na) 700(M + H) 740(M + K) 186 (2-PhCH₂)PhCH₂OPO(Me)Ph C₃₆H₄₂N₃O₈P 675.72 698(M + Na) 674(M − H) 714(M + K)788(M + TFA) 187 (2-PhCH₂)Ph CH₂OPOPh₂ C₄₁H₄₄N₃O₈P 737.79 760(M + Na)736(M − H) 776(M + K) 850(M + TFA) 188 (2-Ph)Ph CH₂OPO(Me)Ph C₄₀H₄₂N₃O₈P661.68 684(M + Na) 660(M − H) 700(M + K) 774(M + TFA) 189 (2-Ph)PhCH₂OPOPh₂ C₃₅H₄₀N₃O₈P 723.75 746(M + Na) 722(M − H) 762(M + K) 836(M +TFA)

EXAMPLE 190

[0330]

[0331] (3S)-3-[N-(N′-(1-Naphthyl)Oxamyl)Homoprolinyl]Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-Oxopentanoic Acid

[0332] Part A: [N-(1-Naphthyl)Oxamyl]Homoproline

[0333] To a solution of N-(1-naphthyl)oxamic acid (0.108 g, 0.50 mmol,see Example 1, Part A) in N-methylpyrrolidone(1.0 mL)-CH₂Cl₂(1.0 mL) atroom temperature under nitrogen was addedO-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophate(0.209 g, 0.55 mmol). After stirring for 20 min, the mixture was treatedwith homoproline methyl ester (0.072 g, 0.50 mmol) anddiisopropylethylamine (0.26 mL, 1.5 mmol). After stirring at roomtemperature for 4 hrs, the mixture was partitioned between EtOAc-water.The organic phase was washed with water, 5% KHSO₄, saturated NaHCO₃ andsaturated NaCl solutions, dried over anhydrous Na₂SO₄ and evaporated togive crude [N-(1-naphthyl)oxamyl]homoproline (0.156 g, 92%) as acolorless glass. TLC (EtOAc-hexane; 1:1) Rf=0.70.

[0334] To a solution of the crude methyl ester (0.156 g, ca 0.46 mmol)in dioxane(0.75 mL)-water(0.25 mL) was added 1.0 N LiOH solution (0.5mL, 0.5 mmol). After stirring at room temperature for 1 hr, the mixturewas acidified with 1.0 N HCl and extracted with EtOAc. The extract waswashed with saturated NaCl solution, dried over anhydrous Na₂SO₄ andevaporated to give the title compound (0.105 g, 70%) as a white solidafter trituration with Et₂O.

[0335] Part B:(3S,4RS)-3-[N-(N′-(1-Naphthyl)Oxamyl)Homoprolinyl]Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-HydroxypenanoicAcid tert-Butyl Ester

[0336] To a solution of [N-(1-naphthyl)oxamyl]homoproline (0.483 g, 1.48mmol) in N-methylpyrrolidone(0.5 mL)-CH₂Cl₂(14 mL) at 0° C. undernitrogen was addedO-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophate(0.676 g, 1.78 mmol). After stirring for 20 min, the mixture was treatedwith a solution of(3S,4RS)-3-amino-5-(2′,3′,5′,6′-tetrafluorophenoxy)-4-hydroxypentanoicacid tert-butyl ester (0.540 g, 1.54 mmol, see Example 49, Part C) inCH₂Cl₂ (4.0 ml,) followed by diisopropylethylamine (0.50 mL, 2.9 mmol).After stirring at 0° C. for 3 hrs and at room temperature for 16 hrs,the mixture was partitioned between EtOAc-water. The organic phase waswashed with water, 5% KHSO₄, saturated NaHCO₃ and saturated NaClsolutions, dried over anhydrous Na₂SO₄ and evaporated to dryness.Purification by flash chromatography on silica gel eluting withEtOAc-hexane (1:2) gave the title compound (0.268 g, 27%) as atan foam.TLC (EtOAc-hexane; 1:1) Rf=0.39.

[0337] Part C:(3S)-3-[N-(N′-(1-Naphthyl)Oxamyl)Homoprolinyl]Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-OxoypentanoicAcid tert-Butyl Ester

[0338] To a solution of(3S,4RS)-3-[N-(N′-(1-naphthyl)oxamyl)homoprolinyl]amino-5-(2′,3′,5′,6′-tetrafluorophenoxy)-4-hydroxypentanoicacid tert-butyl ester (0.251 g, 0.38 mmol) in CH₂Cl₂ (4 mL) at roomtemperature under nitrogen was added Dess-Martin Periodinane (0.201 g,0.475 mmol). After stirring at room temperature for 30 min, the mixturewas diluted with EtOAc, washed with 1.0 M Na₂S₂O₃, saturated NaHCO₃ andsaturated NaCl solutions, dried over anhydrous Na₂SO₄ and evaporated toa dryness. The residue was purified by flash chromatography on silicagel eluting with CH₂Cl₂-Et₂O-hexane (1:2:2) then EtOAc-hexane (1:2) togive the title compound (0.160 g, 64%) as a white foam.TLC(EtOAc-hexane; 1:1) Rf=0.57.

[0339] Part D:(3S,4RS)-3-[N-(N′-(1-Naphthyl)Oxamyl)Homoprolinyl]Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-OxoypentanoicAcid

[0340] To a solution of(3S)-3-[N-(N′-(1-naphthyl)oxamyl)homoprolinyl]amino-5-(2′,3′,5′,6′-tetrafluorophenoxy)-4-oxopentanoicacid tert-butyl ester (0.152 g, 0.23 mmol) in CH₂Cl₂(1.0 mL)-anisole(0.4mL) at room temperature under nitrogen was added trifluoroacetic acid(1.0 mL). The resulting clear solution was stirred at room temperaturefor 1 hr. evaporated to dryness and chased with toluene-CH₂Cl₂ (1:1).The residue was triturated with hexane to give the title compound (0.103g, 74%) as an off-white solid. TLC(MeOH—CH₂Cl₂; 1:9) Rf=0.33. MS(ES) forC₂₉H₂₅F₄N₃O₇ (MW 603.53): positive 626(M+Na); negative 602(M−H).

EXAMPLE 191

[0341]

[0342] (3S)-3-[N-(N′-(1-Naphthyl)Oxamyl)Indoline-2-Carbonyl]Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-Oxopentanoic Acid

[0343] Part A: [N-(1-Naphthyl)Oxamylllndoline-2-Carboxylic Acid EthylEster

[0344] To a solution of N-(1-naphthyl)oxamic acid (2.3 7 g, 11 mmol, seeExample 1, Part A) in N-methylpyrrolidone(7.0 mL)-CH₂Cl₂(40 mL) at 0° C.(ice bath) under nitrogen was added 1,1′-carbonyldiimidazole (1.96 g,12.1 mmol).After stirring at 0° C. for 1.5 hrs and at room temperaturefor 0.5 hrs, (S)indoline-2-carboxylic acid ethyl ester hydrochloride(1.25 g, 5.5 mmol) and diisopropylethylamine (1.1 mL, 6.4 mmol) wasadded. After stirring at room temperature for 18 hrs, the mixture wasdiluted with EtOAc, washed successively with 5% KHSO₄, saturated NaHCO₃and saturated NaCl solutions, dried over anhydrous Na₂SO₄ and evaporatedto dryness. The crude product was purified by flash chromatography onsilica gel eluting with CH₂Cl₂-Et₂O-hexane (1:1:3) to give the titlecompound (0.472 g, 22%) as a pale yellow oil. TLC(CH₂Cl₂-Et₂O-hexane;1:1:3) Rf=0.48.

[0345] Part B:(3S)-3-[N-(N′-(1-Naphthyl)Oxamyl)Indoline-2-Carbonyl]Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-OxopentanoicAcid

[0346] Starting from [N-(1-naphthyl)oxamyl]indoline-2-carboxylic Acidethyl ester, and following the methods described in Example 104, Parts Athrough D, the title compound was also prepared. MS(ES) for C₃₂H₂₃F₄N₃O₇(MW 637.54): positive 660(M+Na),676(M+K); negative 636(M−H),672(M+Cl),750(M+TFA).

EXAMPLE 192

[0347]

[0348] (3S)-3-[N-(N′-(1-Naphthyl)Oxamyl)Cyclohexylglycinyl]Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-Oxopentanoic Acid

[0349] Part A:(3S,4RS)-3-[(N-9-Fluorenylmethoxcycarbony)Cyclohexylpolycinyl]Amnino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-LiydroxypentanoicAcid tert-Butyl Ester

[0350] To a solution of (N-9-fluorenylmethoxycarbonyl) cyclohexylglycine(0.514 g, 1.35 mmol) and(3S,4RS)-3-amino-5-(2′,3′,5′,6′-tetrafluorophenoxy)-4hydroxypentanoicacid tert-butyl ester (0.479 g, 1.36 mmol, see Example 92, Part C) inCH₂Cl₂(10 mL) at 0° C. (ice bath) under nitrogen was addedO-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophate(0.619 g, 1.62 mmol) and diisopropylethylamine (0.47 mL, 2.7 mmol).After stirring at 0° C. for 3 hrs, the reaction was allowed to warm toroom temperature. After stirring at room temperature for 16 hrs, themixture was partitioned between EtOAc-water. The organic phase waswashed with water, 5% KHSO₄, saturated NaHCO₃ and saturated NaClsolutions, dried over anhydrous Na₂SO₄ and evaporated to dryness. Theresidue was purified by flash chromatography eluting with EtOAc-hexane(1:2) to give the title compound (0.481 g, 50%) as a white solid.TLC(EtOAc-hexane; 1:2) Rf=0.42.

[0351] Part B: (3S,4RS)-3-Cyclohexylglycinyl]Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-HydroxypentanoicAcid tert-Butyl Ester

[0352] A solution of(3S,4RS)-3-[(N-9-fluorenylmethoxycarbonyl)cyclohexylglycinyl]amino-5-(2′,3′,5′,6′-tetrafluorophenoxy)-4-hydroxypentanoic acidtert-butyl ester (0.478 g, 0.67 mmol) in piperidine(0.1mL)/dimethylformamide(2.0 mL) was stirred at room temperature undernitrogen for 1 hr. The mixture was diluted with EtOAc, washed with waterand saturated NaCl solution, dried over anhydrous anhydrous Na₂SO₄ andevaporated to dryness. The residue was purified by flash chromatographyeluting with EtOAc-hexane (1:2) to give the title compound (0.121 g,45%) as awhite solid. TLC(MeOH—CH₂Cl₂; 5:95) Rf=0.38.

[0353] Part C:(3S,4RS)-3-[N-(N′-(1-Naphthyl)Oxamyl)Cyclohexylglycinyl]Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-HydroxypentanoicAcid tert-Butyl Ester

[0354] To a solution of N-(1-naphthyl)oxamic acid (0.088 g, 0.41 mmol,see Example 1, Part A) and(3S,4RS)-3-(cyclohexylglycinyl)amino-5-(2′,3′,5′,6′-tetrafluorophenoxy)-4-hydroxypentanoicacid tert-butyl ester (0.110 g, 0.27 mmol) in N-methylpyrrolidone(0.5mL)-CH₂Cl₂(3.0 mL) at 0° C. under nitrogen was addedO-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophate(0.125 g, 0.32 mmol) and diisopropylethylamine (90 μL, 0.54 mmol). Afterstirring at 0° C. for 3 hrs and at room temperature for 16 hrs, themixture was partitioned between EtOAc-water. The organic phase waswashed with water, 5% KHSO₄, saturated NaHCO₃ and saturated NaClsolutions, dried over anhydrous Na₂SO₄ and evaporated to dryness. Theresidue was purified by flash chromatography on silica gel eluting withEtOAc-hexane (1:2) to give the title compound (0.094 g, 50%) as a whitefoam. TLC(EtOAc-hexane; 1:1) Rf=0.50.

[0355] Part D:(3S)-3-[N-(N′-(1-Naphthy)Oxamyl)Cyclohexylglycinyl]Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-OxoypentanoicAcid tert-Butyl Ester

[0356] To a solution of(3S,4RS)-3-[N-(N′-(1-naphthyl)oxamyl)cyclohexylglycinyl]amino-5-(2′,3′,5′,6′-tetrafluorophenoxy)-4-hydroxypentanoic acidtert-butyl ester (0.082 g, 0.12 mmol) in CH₂Cl₂(1 mL)-CH₃CN(2mL)-DMSO(0.2 mL) at room temperature under nitrogen was addedDess-Martin Periodinane (0.145 g, 0.34 mmol). After stirring at roomtemperature for 1 hr, the mixture was diluted with EtOAc, washed with1.0 M Na₂S₂O₃, saturated NaHCO₃ and saturated NaCl solutions, dried overanhydrous Na₂SO₄ and evaporated to a dryness. The residue was purifiedby flash chromatography on silica gel eluting with EtOAc-hexane (1:2then 1:1) to give the title compound (0.068 g, 83%) as a tan foam.TLC(EtOAc-hexane; 1:1) Rf=0.63.

[0357] Part E: (3S,4RS)-3-[N-(N′-(1-Naphthyl)Oxamyl)Cyclohexylglycinyl]Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-OxoypentanoicAcid

[0358] To a solution of(3S)-3-[N-(N′-(1-naphthyl)oxamyl)_cyclohexylglycinyl]amino-5-(2′,3′,5′,6′-tetrafluorophenoxy)-4-oxopentanoic acid tert-butylester (0.065 g, 0.23 mmol) in CH₂Cl₂(1.0 mL)-anisole(0.2 mL) at roomtemperature under nitrogen was added trifluoroacetic acid (1.0 mL). Theresulting clear solution was stirred at room temperature for 30 min,evaporated to dryness and chased with toluene-CH₂Cl₂ (1:1). The residuewas triturated with Et₂O to give the title compound (0.034 g, 56%) as anoff-white solid. TLC(MeOH—AcOH—CH₂Cl₂; 1:1:32) Rf=0.45. MS(ES) forC₃₁H₂₉F₄N₃O₇ (MW 631.58): positive 654(M+Na); negative 630(M−H).

EXAMPLES 193-200

[0359] Starting from(3S,4RS)-3-amino-5-(2′,3′,5′,6′-tetrafluorophenoxy)-4-entanoicjydroxy-pentanoic acid tert-butyl ester (see Example 178, Part C) andfollowing the general methods described in Example 192, Parts A throughE, the compounds shown below in Table 10 were also prepared: TABLE 10

MS(ES) Ex. A Formula MW pos. neg. 193 norleucine C₂₉H₂₇F₄N₃O₇ 605.54628(M + Na) 604(M − H) 644(M + K) 640(M + Cl) 718(M + TFA) 194(t-butyl)glycine C₂₉H₂₇F₄N₃O₇ 605.54 606(M + H) 604(M − H) 628(M + Na)640(M + Cl) 644(M + K) 718(M + TFA) 195 (t-butyl)alanine C₂₀H₂₉F₄N₃O₇619.57 620(M + H) 618(M − H) 642(M + Na) 732(M + TFA) 658(M + K) 196phenylglycine C₃₁H₂₃F₄N₃O₇ 625.53 626(M + H) 624(M − H) 648(M + Na)660(M + Cl) 664(M + K) 738(M + TFA) 197 phenylalanine C₃₂H₂₅F₄N₃O₇639.56 640(M + H) 638(M − H) 662(M + Na) 674(M + Cl) 678(M + K) 712(M +TFA) 198 homophenylalanine C₃₃H₂₇F₄N₃O₇ 653.59 654(M + H) 652(M − H)676(M + Na) 688(M + Cl) 692(M + K) 766(M + TFA) 199 1-aminocyclopentaneC₂₉H₂₅F₄N₃O₇ 603.53 626(M + Na) 602(M − H) carboxylic acid 642(M + K)200 histidine C₂₉H₂₃F₄N₅O₇ 629.15 630(M + H) 28(M − H)

EXAMPLE 201

[0360]

[0361] (3S)-3-[N-(N′-(1-Naphyhyl)Oxamyl)Methioninyl(Sulfoxide)]Amino-5-(2′,3′,5′,6′-Tetrafluoropenoxy)-4-Oxopentanoic Acid

[0362] Part A:(3S,4RS)-3-[N-(N′-(1-Naphthyl)Oxamyl)Methioninyl]Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-HydroxypentanoicAcid tert-Butyl Ester

[0363] Starting from (N-9-fluorenylmethoxycarbonyl) methionine andfollowing the methods described in Example 106, Parts A through C, thetitle compound was also prepared. TLC([tOAc-hexane; 1:2) Rf=0.39.

[0364] Part B;(3S)-3-[N-(N′-(1-Naphthyl)Oxamyl)Methioninyl(Sulfoxide)]Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-OxoypentanoicAcid tert-Butyl Ester

[0365] To a solution of(3S,4RS)-3-N-(N′-(1-naphthyl)oxamyl)methioninyl]amino-5-(2′,3′,5′,6′-tetrafluorophenoxy)-4-hydroxypentanoicacid tert-butyl ester (0.251 g, 0.37 mmol) in CH₂Cl₂(4.0 mL) at roomtemperature under nitrogen was added Dess-Martin Periodinane (0.203 g,0.48 mmol). After stirring at room temperature for 1 hr. the mixture wasdiluted with EtOAc, washed with 1.0 M Na₂S₂O₃, saturated NaHCO₃ andsaturated NaCl solutions, dried over anhydrous Na₂SO₄ and evaporated toa dryness. The residue was purified by flash chromatography on silicagel eluting with EtOAc-hexane (1:2 then 1:1) followed by MeOH—CH₂Cl₂(5:95 then 1:9) to give a mixture of two isomeric sulfoxides (0.225 g);TLC(MeOH—CH₂Cl₂; 1:9) Rfs 0.48 and 0.43. The mixture wasre-chromatographed on silica gel eluting with isopropanol-CH₂Cl₂ (2.5%to 5% to 10%) to give sulfoxide isomer A (less polar, 0.051 g),sulfoxide isomer B (more polar, 0.086 g) and a mixture of isomers A andB (0.040 g). Both isomers have virtually identical mass spectra. MS(ES)for C₃₂H₃₃F₄N₃O₈S (MW 695.68): positive 718(M+Na); negative 694(M−H).

[0366] Part C:(3S,4RS)-3-[N-(N′-(1-Naphthy)Oxamyl)Methioninyl(Sulfoxide)]Amino-5-(2′,3′,5′,6′-Tetrafluorophenoxy)-4-Oxoypentanoic Acid

[0367] To a solution of(3S)-3-[N-(N′-(1-naphthyl)oxamyl)methioninyl(sulfoxide)]amino-5-(2′,3′,5′,6′-tetrafluorophenoxy)-4-oxopentanoic acid tert-butylester (isomer A, 0.046 g, 0.07 mmol) in CH₂Cl₂(2.0 mL)-anisole(0.1 mL)at room temperature under nitrogen was added trifluoroacetic acid (1.0mL). The resulting clear solution was stirred at room temperature for 30min, evaporated to dryness and chased with toluene-CH₂Cl₂ (1:1). Theresidue was triturated with Et₂O-hexane to give the title compound,isomer A (0.034 g, 81%) as an off-white solid. TLC(MeOH—AcOH—CH₂Cl₂;1:1:32) Rf=0.20. MS(ES) for C₂₈H₂₅F₄N₃O₈S (MW 639.57): positive640(M+H),662(M+Na),678(M+K); negative 638(M−H), 752(M+TFA). Under thesame conditions sulfoxide isomer B (0.081 g, 0.12 mmol) gave the titlecompound, isomer B (0.055 g, 74%). MS(ES) for C₂8H₂₅F₄N₃O₈S (MW 639.57):positive 640(M+H),662(M+Na),678(M+K); negative 63 8(M−H), 674(M+Cl),752(M+TFA).

EXAMPLE 202

[0368]

[0369] (3S)-3- [N-(N′-(1-Naphthyl)Oxamyl)Homoprolinyl]Amino-4-Oxobutanoic Acid

[0370] Part A:(3S)-3-[N-(N′-(1-Naphthyl)Oxamyl)Homoprolinyl]Amino-4-Oxobutanoic Acid(tert)-Butyl Ester Semicarbazone

[0371] To a solution of [N-(1-naphthyl)oxamyl]homoproline (0.103 g, 0.32mmol, see Example 104, Part A) in CH₂Cl₂ (3.0 mL) at 0° C. undernitrogen was added was added hydroxybenzotriazole hydrate (0.058 g)followed by 1-ethyl-3-(3′,3′-dimethyl-1′-aminopropyl)carbodiimidehydrochloride (0.91 g, 0.47 mmol). After stirring at 0° C. for 10 min,the mixture was treated with (3S)-amino-4-oxobutanoic acid (tert)-butylester semicarbazone, p-toluenesulfonate salt (0.127 g, 0.32 mmol) andN-methylmorpholine (42 liL, 0.38 mmol). After stirring at 0° C. for 2hrs, the mixture was concentrated and the residue partitioned betweenEtOAc-5% KHSO₄. The organic phase was washed with 5% KHSO₄, saturatedNaHCO₃ and saturated NaCl solutions, dried over anhydrous Na₂SO₄ andevaporated to give the crude title compound (0.119 g, 70%) as acolorless glass.

[0372] Part B:(3S)-3-[N-(N′-(1-Naphthyl)Oxamyl)Homoprolinyl]Amino-4-Oxobutanoic AcidSemicarbazone

[0373] To a solution of(3S)-3-[N-(N′-(1-naphthyl)oxamyl)homoprolinyl]amino-4-oxobutanoic acidsemicarbazone tert-butyl ester (0.119 g, 0.21 mmol) in CH2Cl2(2.0mL)-anisole(0.05 mL)-water(0.05 mL) at room temperature under nitrogenwas added trifluoroacetic acid (0.32 mL). The resulting clear solutionwas stirred at room temperature for 18 hrs, evaporated to dryness andchased with toluene-CH2Cl2 (1:1). The residue was triturated with Et2Oto give the title compound (0.079 g, 74%) as a white solid.

[0374] Part C:(3S)-3-[N-(N′-(1-Naphthyl)Oxamyl)Homoprolinyl]Amino-4-Oxobutanoic Acid

[0375] A suspension of(3S)-3-[N-(N′-(1-naphthyl)oxamyl)homoprolinyl]amino-4-oxobutanoic acidsemicarbazone (0.079 g, 0.16 mmol) in 37% aqueous formaldehyde(0.6mL)-acetic acid(0.6 mL)-methanol(1.8 mL) was stirred at room temperatureunder nitrogen for 18 hrs. The resulting clear solution was diluted withwater and mixture concentrated on a rotovap. The aqueous solution wasthen frozen and lyophilized. The residue was taken Up in methanol,filtered through Celite and filtrate evaporated to dryness. Triturationof the residue with Et₂O gave the title compound (0.037 g, 53%) as awhite solid. MS(ES) for C₂₂H₂₃N₃O₆ (MW 425.44): positive 448(M+Na);negative 424(M−H).

EXAMPLE 203

[0376]

[0377] (3S)-3-[N-(N′-(2-(1H-Tetrazol-5-yl)Phenyl)Oxamyl)Valinyl]Amino-4-Oxobutanoic Acid

[0378] Part A: 2-(1′-Phenylmethyl-5′-Tetrazolyl)Aniline Hydrochloride

[0379] A solution of 2-cyano-acetanilide (0.801 g, 5.0 mmol) andtri-n-butyltin azide (2.05 mL, 7.5 mmol) in anhydrous toluene (10 mL)was heated at reflux for 48 hrs. The mixture was allowed to cool to roomtemperature and treated with 2.0 N HCl in Et₂O (5.0 mL). The resultingprecipate was collected by suction, washed with hexane and dried invacuo to give 2-(lH-tetrazol-5-yl)acetanilide (0.917 g, 90%) as a whitesolid.

[0380] To a suspension of 2-(1H-tetrazol-5-yl)acetanilide (0.203 g, 1.0mmol) in tetrahydrofuran (2.0 mL) at 0° C. under nitrogen was addedtriethylamine (0.170 mL, 1.2 mmol) and benzyl bromide (0.125 mL, 1.05mmol). After stirring at 0° C. for 3 hrs and at room temperature for 16hrs, the mixture was partitioned between EtOAc-water. The organic phasewas washed with saturated NaHCO₃ and saturated NaCl solutions, driedover anhydrous Na₂SO₄ and evaporated to a dryness. The residue wastriturated with hexane to give2-(1′-phenylmethyl-5′-tetrazolyl)acetanilide (0.218 g, 74%) as a whitesolid. ¹H—NMR indicates that the product is a single regioisomer.Assignment of regiochemistry should be considered tentative.¹H—NMR(CDCl₃): δ 2.22 ppm (3H,s), 5.84 (2H,s), 7.16 (1H, dt, J=7.8, 1.5Hz), 7.40 (6H, m), 8.19 (1H, dd, J 7.8, 1.5 Hz), 8.63 (1H, d, J=8.4 Hz).10.58 (1H, bs).

[0381] A mixture of 2-(1′-phenylmethyl-5′-tetrazolyl)acetanilide (0.216g, 0.74 mmol) and 10% aqueous HCl (3.0 mL) was refluxed for 18 hrs. Themixture was evaporated to dryness and the residue triturated with Et₂Oto give the title compound (0.187 g, 88%) as awhite solid.

[0382] Part B: N-[2-(1′-Phenylmethyl-5′-Tetrazolyl)Phenyl]Oxamic Acid

[0383] To a solution of 2-(1′-phenylmethyl-5′-tetrazolyl)anilinehydrochloride (0.177 g, 0.615 mmol), 4-dimethylaminopyridine (0.008 g,0.065 mmol) and triethylamine (0.19 mL, 1.4 mmol) in CH₂Cl₂ (1.0 mL) at0° C. (ice bath) under nitrogen was added methyl oxalyl chloride (62 μL,0.67 mmol). After stirring at 0° C for 2 hrs, the mixture was allowed tocome to room temperature, stirred for 18 hrs and then partitionedbetween EtOAc-5% KHSO₄. The organic phase was washed with saturated NaClsolution, dried over anhydrous Na₂SO₄ and evaporated to dryness.

[0384] The crude methyl ester (0.207 g, ca 0.615 mmol) was taken up indioxane (2.0 mL) and treated with 1.0 N LiOH solution (0.68 mL, 0.68mmol) and stirred at room temperature for 1 hr. The mixture wasacidified with 1.0 N HCl and extracted with EtOAc. The extract waswashed with saturated NaCl solution, dried over anhydrous Na₂SO₄ andevaporated to dryness. Trituration of the crude product with hexane gavethe title compound (0.121 g, 61%) as awhite solid.

[0385] Part C: (3S)-3-[N-(N′-(2-(1′-Phenylmethyl-5′-Tetrazolyl)Phenyl)Oxamyl)Valinyl]Amino-4-Oxobutanoic Acid Semicarbazone tert-Butyl Ester

[0386] To a solution ofN-[2-(1′-phenylmethyl-5′-tetrazolyl)phenyl]oxamic acid (0.065 g, 0.20mmol) in CH₂Cl₂(2.0 mL) at 0° C. under nitrogen was addedhydroxybenzotriazole hydrate (0.037 g) followed by1-ethyl-3-(3′,3′-dimethyl-1′-aminopropyl)-carbodiimide hydrochloride(0.058 g, 0.30 mmol). After stirring at 0° C. for 10 min, the mixturewas treated with (3S)-3-(valinyl)amino-4-oxobutanoic acid (tert)-butylester semicarbazone (0.066 g, 0.20 mmol, prepared by the methoddescribed for the corresponding leucine analogue in Example 1, Parts Band C) and N-methylmorpholine (26 μL, 0.24 mmol). After stirring at roomtemperature for 16 hrs, the mixture was partitioned between EtOAc-water.The organic phase was washed with water, 5% KHSO₄, saturated NaHCO₃ andsaturated NaCl solutions, dried over anhydrous Na₂SO₄ and evaporated togive the crude title compound (0.090 g, 62%) as a colorless glass.

[0387] Part D: (3S)-3-[N-(N′-(2-(1′H-5′-Tetrazolyl)Phenyl)Oxamyl)Valinyl]Amino-4-OxobutanoicAcid Semicarbazone tert-Butyl Ester

[0388] To a solution of crude(3S)-3-[N-(N′-(2-(1′-phenylmethyl-5′-tetrazolyl)phenyl)oxamyl)valinyl]amino-4-oxobutanoic acid semicarbazone tert-butyl ester(0.089 g, ca.0.14 mmol) in MeOH (1.0 mL) was added 10% Pd-C (0.009 g)and resulting mixture stirred under a hydrogen atmosphere (balloon) for48 hrs. The mixture was filtered through Celite washing the filter cakewith CH₂Cl₂ and the combined filtrates evaporated to dryness. Theresidue was triturated with Et₂O to give the title product (0.060 g,79%) as a white solid.

[0389] Part E:(3S)-3-[N-(N′-(2-(1′H-5′-Tetrazolyl)Phenyl)Oxamyl)Valinyl]Amino-4-OxobutanoicAcid Semicarbazone

[0390] To a solution of(3S)-3-[N-(N′-(2-(1′H-5′-tetrazolyl)phenyl)oxamyl)valinyl]amino-4-oxobutanoic acid tert-butyl ester (0.058, 0.11 mmol) inCH₂Cl₂(1.0 mL)-anisole(0.05 mL) at room temperature under nitrogen wasadded 6.0 M HCl/AcOH (1.0 mL). The resulting solution was stirred atroom temperature for 18 hrs, evaporated to dryness and chased withtoluene-CH₂Cl₂ (1:1). The residue was triturated with Et₂O to give thetitle compound (0.048 g, 92%) as a white solid.

[0391] Part F:(3S)-3-[N-(N′-(2-(1′H-5′-Tetrazolyl)Phenyl)Oxamyl)Valinyl]Amino-4-OxobutanoicAcid

[0392] A solution of(3S)-3-[N-(N′-(2-(1′H-5′-tetrazolyl)phenyl)oxamyl)valinyl]amino-4-oxobutanoic acid semicarbazone (0.048 g, 0.10 mmol) in 37%aqueous formaldehyde(0.4 mL)-acetic acid(0.4 mL)-methanol(1.2 mL) wasstirred at room temperature under nitrogen for 18 hrs. The resultingclear solution was diluted with water and mixture concentrated on arototvap. The aqueous solution was then frozen and lyophilized. Theresidue was taken up in methanol, filtered through Celite and filtrateevaporated to dryness. Trituration of the residue with Et₂O gave thetitle compound (0.025 g, 59%) as a white solid. MS(ES) for C₁₈H₂₁N₇O₆(MW 431.41): positive 454(M+Na); negative 430(M−H).

EXAMPLE 204

[0393]

[0394] (3S)-3-[N-(N′-(1-Adamantanyl)Oxamyl)Valinyl] Amino-4-OxobutanoicAcid

[0395] Part A:(3S)-3-[N-(9-Fluorenylmethoxycabonyl)Valinyl]Amino-4-Oxobutanoic Acid(tert-Butyl) Ester Semicarbazonyl-4-[2′-(4-Ethyl-Phenoxyacetyl)]Aminomethylpolystrene

[0396] Aminomethylpolystryene resin (10.0 g, 100-200 mesh, 0.71 meq/g)was placed in a 200 mL filter tube equipped with a vacuum stopcock andglass frit and washed successively with CH₂Cl₂(50mL)/dimethylformamide(50 mL), diisopropylethylamine(5mL)/dimethylformamide(30 mL), dimethylformamide (2×50 mL) andtetrahydrofuran (30 mL). The resin was suspended in tetrahydrofuran(20mL)/N-methylpyrrolidone(20 mL) with nitrogen agitation through thebottom of the frit and treated with diisopropylethylamine (1.9 ML, 10.9mmol) and (3S)-3-(9-fluorenylmethoxycabonyl)amino-4-oxobutanoic acid(tert-butyl) ester semicarbazonyl-4-[2′-(4-ethyl-phenoxyacetic acid)](2.24 g, 3.56 mmol). After all of the solid had dissolved (approx. 10min), the mixture was treated with pyBOP[benzotriazolyloxy-tris(N-pyrolidinyl)phosphonium hexafluorophosphate,2.78 g, 5.34 mmol) in one portion. After mixing by nitrogen agitationfor 3 hrs, the supernatant was removed by suction and the resin washedsuccessively with tetrahydrofuran (2×50 mL), dimethylformamide (3×50 mL)and CH₂Cl₂ (2×50 mL). Unreacted amine groups were capped by treatmentwith a rmixture of acetic anhydride(10 mL)/dimethylformamide(30mL)/diisopropylethylamine(1.0 mL). After mixing by nitrogen agitationfor 1 hr, the supernatant was removed by suction and the resin washedwith dimethylformamide(4×50 mL).

[0397] The resin was treated with piperidine(10 mL)/dimethylformamide(40mL) and mixed by nitrogen agitation for 1 hr. The supernatant wasremoved by suction and the resin washed with dimethylformamide(4×50 mL)and tetrahydrofuran (50 mL).

[0398] The resin was suspended in tetrahydrofuran(20mL)/N-methylpyrrolidone(20 mL), treated withN-(9-fluorenylmethoxycabonyl)valine (3.63 g, 10.7 mmol),diisopropylethylamine (5.7 miL, 32.7 mmol) and pyBOP (8.34 g, 16.0 mmol)and mixed by nitrogen agitation for 2.5 hrs. The supernatant was removedby suction and the resin washed successively with dimethylformamide(3×40 mL) and CH₂Cl₂ (3×40 mL), methanol (2×40 mL) and Et₂O (2×40 mL).The resin was dried in vacuo to give the title product (12.69 g,quantitative). Based on the starting semicarbazone-acid, the resinloading was calculated as approximately 0.28 meq/g.

[0399] Part B:(3S)-3-[N-(N′-(1-Adamantanyl)Oxamyl)Valinyl]Amino-4-Oxobutanoic Acid

[0400] An aliquot of the Part Aresin (0.125 g, ca 0.035 mmol) was placedin a 6 mL Supelco™ filtration tube equipped with a 20 μm polyethylenefrit, treated with piperidine-dimethylformamide (1.0 mL, 1:4 v/v) andmixed on an orbital shaker for 1 hr. The supernatant was removed bysuction and the resin washed with dimethylformamide (4×1.0 mL) andCH₂Cl₂ (3×1.0 mL). The resin was treated with 0.5M iPr₂NEt inN-methylpyrrolidone (0.40 mL, 0.20 mmol), (1-adamantanyl)oxamic acid(0.0246 g, 0.11 mmol) and 0.25MO-(7-azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluronium hexafluorophatein N-methylpyrrolidone (0.40 mL, 0.10 mmol). The mixture was mixed on anorbital shaker under an nitrogen atmosphere for 16 hrs. The supernatantwas removed by suction and the resin washed successively withdimethylformamide (3×1.0 mL) and CH₂Cl₂ (3×1.0 mL), methanol (2×1.0 mL)and Et₂O (2×1.0 mL).

[0401] The resin was treated with 1.0 mL of CH₂Cl₂ and allowed tore-swell for 15 min. The solvent was removed by suction and the resintreated with trifluoroacetic acid-CH₂Cl₂-anisole (1.0 mL, 4:3:1 v/v/v).After mixing on an orbital shaker under nitrogen for 5.5 hrs, thesupernatant was removed by suction and the resin washed with CH₂Cl₂(4×1.0 mL). The resin was treated with 37% aqueous formaldehyde-aceticacid-tetrahydrofuran-trifluoroacetic acid (1.0 mL, 1:1:5:0.025 v/v/v/v)and mixed on an orbital shaker under nitrogen for 4.5 hrs. Thesupernatant was collected by suction, the resin washed withtetrahydrofuran (3×0.5 mL). The combined filtrates were blown down undernitrogen. The residue was taken up in methanol (0.5 mL), filtered andapplied directly to a 3 mL Supelco™ LC-18 reverse phase extraction tubewhich had been pre-conditioned with water, and eluted successively with3 mL each of 10% MeOH-water, 30% MeOH-water, 60% MeOH-water and 90%MeOH-water. The product-containing frac(tions (TLC) were combined andevaporated to dryness to give the title compound (0.0114 g, 77%) as acolorless glass. TLC(AcOH-MeOH-CH₂Cl₂; 1:1:20) Rf=0.23. MS(ES) forC₂₁H₃₁N₃O₆ (MW 421.49): positive 444(M+Na), 460(M+K); negative 420(M−H),534(M+TFA).

EXAMPLES 205-219

[0402] Starting with(3S)-3-[N-(9-fluorenylmethoxycabonyl)valinyl]amino-4-oxobutanoic acid(tert-butyl) ester semicarbazonyl-4-[2′-(4-ethyl-phenoxyacetyl)]aminomethylpolystrene (see Example 204, Part A) and following themethods described in Example 204, Part B, the compounds shown below inTable 11 were also prepared: TABLE 11

MS(ES) Ex. R¹ Formula MW pos. neg. 205 Ph C₁₇H₂₁N₃O₆ 363.37 386(M + Na)362(M − H) 402(M + K) 206 PhCH₂ C₁₈H₂₃N₃O₆ 377.40 400(M + Na) 376(M − H)207 Ph(CH₂)₂ C₁₉H₂₅N₃O₆ 391.42 414(M + Na) 390(M − H) 430(M + K) 504(M +TFA) 208 (2-CF₃)Ph C₁₈H₂₀F₃N₃O₆ 431.37 454(M + Na) 430(M − H) 209(2-t-Bu)Ph C₂₁H₂₉N₃O₆ 419.48 442(M + Na) 418(M − H) 458(M + K) 532(M +TFA) 210 (2-Ph)Ph C₂₃H₂₅N₃O₆ 439.47 462(M + Na) 438(M − H) 478(M + K)552(M + TFA) 211 (2-PhCH₂)Ph C₂₄H₂₇N₃O₆ 453.49 476(M + Na) 452(M − H)492(M + K) 566(M + TFA) 212 (2-PhO)Ph C₂₃H₂₅N₃O₇ 455.47 478(M + Na)454(M − H) 494(M + K) 568(M + TFA) 213 2-naphthyl C₂₁H₂₃N₃O₆ 413.43436(M + Na) 412(M − H) 452(M + K) 526(M + TFA) 214 1-naphthyl C₂₁H₂₃N₃O₆413.43 436(M + Na) 412(M − H) 452(M + K) 526(M + TFA) 2154-Cl-1-naphthyl C₂₁H₂₂ClN₃O₆ 447.87 470/472(M + Na) 446/448(M − H)486/488(M + K) 216 5,6,7,8-tetrahydro-1-naphthyl C₂₁H₂₇N₃O₆ 417.46440(M + Na) 416(M − H) 456(M + K) 530(M + TFA) 2171,2,3,4-tetrahydro-1-naphthyl C₂₁H₂₇N₃O₆ 417.46 440(M + Na) 416(M − H)456(M + K) 530(M + TFA) 218 (1-naphthyl)CH₂ C₂₂H₂₅N₃O₆ 427.46 450(M +Na) 426(M − H) 466(M + K) 540(M + TFA) 219 2-benzimidazoyl C₁₈H₂₁N₅O₆403.39 — —

EXAMPLE 220

[0403]

[0404]N-(5-Acethylamino-2-Tert-Butyl-Phenyl)-N′-[1-(2-Hydroxy-5-Oxo-Tetrahydro-Furan-3-Ylcarbamoyl)-Ethyl]-Oxalamide

[0405] Part A: 2-tert-Butyl-5-nitro-phenylamine

[0406] To H₂SO₄ (50.0 g, 509.79 mmol) was slowly added2-terf-Butylaniline (5.0 g, 33.50 mmol). The mixture was stirred at roomtemperature until it became homogeneous, then was cooled to −10° C.before slow addition of KNO₃ in small portions (5.00 g, 49.45 mmol) viaa powder addition funnel. After stirring at −10° C. for an hour, thereaction mixture was poured over a small portion of ice in a 250 mLbeaker, allowed to stand for 10 min, then filtered and discarded whiteprecipitate. The aqueous solution was neutralized with NH₄0H until pH8-9 (Litmus pH paper) and partitioned between EtOAc/water. The organicphase was washed with water, and saturated NaCt solutions, dried overanhydrous Na₂SO₄ and evaporated to a dryness (brown oil). The brownresidue was recrystallized from methanol/water to give the desiredcompound (4.1 g, 63%) as dark brown crystals. TLC (20% EtOAc/Hexane)R_(f)=0.66; ¹HNMR (CDCl₃) δ 7.55 (dd, J=8.4, 2.4 Hz, 1H), 7.46 (d, J=2.7Hz, 1H), 7.35-7.33 (d, J=8.7 Hz), 4.16 (br.s, 21H), 1.44 (s, 9H).

[0407] Part B: N-(2-tert-Butyl-5-nitro-phenyl)-oxalamic acid methylester

[0408] To a suspension of 2-tert-Butyl-5-nitro-phenylamine (4.04 g,20.80 mmol) in CH₂Cl₂ (20 mL) at 0° C. was added 1.1 equiv of Methylchlorooxoacetate (2.10 mL, 22.88 mmol) and followed by dropwise additionof 1.1 equiv of Et₃N (3.19 mL, 22.88 mmol). After stirring at 0° C. andallowing to warm up to room temperature over 4 hrs, the reaction mixturewas treated with water (20 mL), stirred for 10 min and then partitionedbetween EtOAc/water. The organic phase was washed with 0.5N HCl (2×25mL), and saturated NaCl solution (50 mL), dried over Na₂SO₄ andevaporated to dryness. The residue was purified by flask columnchromatography on silica gel eluting with EtOAc/Hexane (5-25%) to givethe title compound (3.25 g, 56%) as light yellow crystals. TLC (30%EtOAc/Hexane) R_(f)=0.39; ¹HNMR (CDCl₃) δ 9.36 (br.s, 1IH), 8.97 (d,J=2.4 Hz, 1H), 8.02 (dd, J=9.0, 2.4 Hz, 1H), 7.60 (d, J=9.0 Hz, 1H),4.03 (s, 3H), 1.52 (s, 9H).

[0409] Part C: N-(5-Amino-2-tert-butyl-phenyl)-oxalamic acid methylester

[0410] To a suspension of N-(2-tert-Butyl-5-nitro-phenyl)-oxalamic acidmethyl ester (1.50 g, 5.32 mmol) in MeOH (100 mL) was added Pd/C 10 mol%(0.10 g). The reaction flask was purged with H₂ (1 atm)/vacuum threetimes, then stirred under H₂ (1 atm) at room temperature. After stirringfor 45 min, the reaction was filtered through celite and evaporated todryness. The residue was triturated with hexane to give the titlecompound (1. 12 g, 84%) as a gray solid. TLC (40% EtOAc/Hexane)R_(f)=0.21; ¹HNMR (CDCl₃) δ 9.21 (br.s, 1H), 7.47 (d, J=2.4 Hz, IH),7.18 (d, J=8.7 Hz, IH), 6.50 (dd, J=8.4, 2.7 Hz, 1H), 4.00 (s, 3H), 1.42(s, 9H); MS(ES) for C13H18N205 (MW=250.29) positive 251 (MH+).

[0411] Part D: N-(5-Acetylamino-2-tert-butyl-phenyl)-oxalamic acidmethyl ester

[0412] To a suspension of N-(5-Amino-2-tert-butyl-phenyl)-oxalamic acidmethyl ester (1.10 g, 4.39 mmol) in pyridine (5 mL) at room temperaturewas added 1.1 equiv of Acetic anhydride (0.45 mL, 4.83 mmol). Afterstirring at room temperature for 30 min, the reaction mixture wastreated with 0.5N CuSO₄ (50 mL), stirred for 5 min and then partitionedbetween EtOAc/water. The organic phase was washed with 0.5N HCl (2×25mL), and saturated NaCl solution (50 mL), dried over Na₂SO₄ andevaporated to dryness to give the tittle compound as a white foam (1.28g, >98%). TLC (70% EtOAc/Hlexane) R_(f)=0.43; ¹HNMR (CDCl₃) δ 9.26(br.s, 1H), 7.90 (d, J=2.4 Hz, 1H), 7.65-7.61 (dd, J=8.7, 2.4 Hz, 1H),7.47 (br.s, 1H), 7.35 (d, J=8.7 Hz, 1H), 4.10 (s, 3H), 2.16 (s, 3H),1.44 (s, 9H); MS(ES) for C₁₅H₂₀N₂O₄ (MW=292.33): positive 293 (MH+).

[0413] Part E: N-(5-Amino-2-tert-butyl-phenyl)-oxalamic acid

[0414] To a suspension of N-(5-Acetylamino-2-tert-butyl-pbenyl)-oxalamicacid methyl ester (1.28 g, 4.38 mmol) in 1,4-Dioxane (5 mL) at roomtemperature was added 1.05 equiv of 1.ON LiOH (4.60 niL, 4.60 mmol).After stirring at room temperature for 30 min, the reaction mixture wastreated with 0.5N HCl (20 mL), stirred for 5 min and then partitionedbetween EtOAc/water. The organic phase was washed with saturated NaClsolution (50 mL), dried over Na₂SO₄ and evaporated to dryness. The oilyresidue was recrystallized from CH₂Cl₂/Hexane to give the title compoundas a white fine crystal (1.74 g, 96%). ¹HNMR (DMSO-d₆) δ 10.05 (s, 1H),9.96 (s, 1H), 7.50 (d, J=2.1 Hz, 1IH), 7.42 (dd, J=9.0, 2.1 Hz, 1H),7.33 (d, J=8.7 Hz, 1H), 2.02 (s, 3H), 1.30 (s, 9H); MS(ES) forC₁₄H₁₈N₂O₄ (MW=278.30): negative 277 ([M−H]⁻).

[0415] Part F: (2S)-2-[(5-Acetylamino-2-tert-butyl-phenylaminooxalyl)-amino]-propaneperoxoicacid methyl ester

[0416] To a solution of N-(5-Amino-2-tert-butyl-phenyl)-oxalamic acid(0.35 g, 1.36 mmol) in CH₂Cl₂/1-methyl-2-pyrolidinone (NMW) (1:1) (3 mL)at room temperature under N₂ was added 1.5 equivO-(7-Azabenzotriazol-1-yl)-N,N,N′,N′-tetramethyluroniumhexafluorophosphate (HATU) (0.72 g, 1.89 mmol). The mixture solution wasstirred at room temperature under N₂ for 45 min before addition ofHClH-Ala-OMe (0.16 g, 1.51 mmol), followed by Et₃N (0.53 mL, 3.78 mmol).After stirring at room temperature for 2 hrs, the mixture waspartitioned between EtOAc-water. The organic phase was washed withwater, 5% KHSO₄, saturated NaHCO₃, and saturated NaCl solutions, driedover anhydrous Na₂SO₄ and evaporated to give crude title compound. Theresidue was purified by flask column chromatography on silica geleluting with EtOAc/Hexane (20-60%) to give the tittle compound (0.34 g,74%) as a white solid. TLC (80% EtOAc/Hexane) R_(f)=0.51; ¹HNMR (CDCl₃)δ 9.63 (br.s, 1H), 8.11 (d, J=7.8 Hz, 1H), 8.01 (d, J=2,4 Hz, 1H), 7.60(dd, J=8.4, 2.1 Hz, 1H), 7.50 (s, 1H), 7.35 (d, J=98.7 Hz, lH),4.71-4.61 (dq, J=15, 7.5 Hz, 1H), 3.80 (s, 3H), 2.15 (s, 3H), 1.55 (d,J=7.2 Hz, 3H), 1.44 (s, 9H). MS(ES) for C₁₈H₂₅N₃O₆ (MW=363.41): positive381 ([M+NH₄]⁺).

[0417] Part G:(2S)-2-[(5-Acetylamino-2-tert-butyl-phenylaminooxalyl)-amino]-propionicacid

[0418] To a suspension of(2S)-2-[(5-Acetylamino-2-tert-butyl-phenylaminooxalyl)-amino]-propionicacid methyl ester (0.32 g, 0.88 mmol) in 1,4-Dioxane (2 mL) at roomtemperature was added 1.05 equiv of 1.ON LiOH (0.92 mL, 0.92 mmol).After stirring at room temperature for 30 min, the reaction mixture wastreated with 0.5N HCl (10 mL), stirred for 5 min and then partitionedbetween EtOAc/water. The organic phase was washed with saturated NaClsolution (50 mL), dried over Na₂SO₄ and evaporated to dryness. The oilresidue was recrystallized from EtOAc/Hexane to give the title compoundas a white fine powder (0.31 g, 99%). ¹HNMR (CDCl₃) δ 9.66 (s, 1H), 8.42(d, J=8.4 Hz, 1H), 8.14 (s, 1H), 7.91 (s, 1H), 7.55-7.52 (dd, J=8.4, 1.8Hz, 1H), 7.33 (d, J=9.0Hz, 1H), 4.72-4.63 (q, J=7.5 Hz, 1H), 4.20-3.90(bs, 1H), 2.13 (s, 3H), 1.53 (d, J=7.2 Hz, 3H), 1.41 (s, 3H); MS(ES) forC₁₄H₁₈N₂O₄ (MW=349.38): negative 348 ([M−H]⁻).

[0419] Part H: 3-Benzyloxvcarbonylamino-N-methoxy-N-methyl-succinamicacid tert-butyl ester

[0420] To a solution of 2-Benzyloxycarbonylamino-succinic acid4-tert-butyl ester (15.00 g, 46.39 mmol) in CH₂Cl₂ (150 mL) at 0° C.under N₂ was added 1.2 equiv of1-(3-Dimethylaminopropyl)-3-ethylcarbodiimideHCl (EDAC) (10.67 g, 55.67mmol) and 1.1 equiv of HOBt.H₂O (7.81 g, 51.03 mmol). The mixture wasstirred at 0° C. under N₂ for 15 min before addition of HCl HN(OMe)Me(5.88g, 60.31 mmol), followed by 4-Methylmorpholine (NMM) (7.65 mL,69.59 mmol). After stirring at 0° C. to room temperature for 18hrs, themixture was partitioned between EtOAc/water. The organic phase waswashed with water, 5% KHSO₄, saturated NaHCO₃, and saturated NaClsolutions, dried over anhydrous Na₂SO₄ and evaporated to give crudetitle compound. The residue was recrystallized from Et₂O/n-Hexane togive the tittle compound (14.61 g, 86%) as a white solid. TLC (50%EtOAc/Hexane) R_(f)=0.58; ¹HNMR (CDCl₃) δ 7.34 (m, 5H), 5.64 (d, J=9.3Hz, 1H), 5.15-5.05 (dd, J=18.3, 12.3 Hz, 2H), 5.05-5.00 (m, 1H), 3.78(s, 3H), 3.22 (s, 1H), 2.74-2.68 (dd, J=15.0, 5.4 Hz, 1H), 2.58-2.51(dd, J=15.6, 7.2 Hz, 1H, 1.42 (s, 9H). MS(ES) for C₁₈H₂₆N₂O₆ (MW=366.4):positive 367 (MH⁺).

[0421] Part I: 3-Benzyloxycarbonylamino-4-oxo-butyric acid tert-butylester

[0422] To a suspension of3-Benzyloxycarbonylamino-N-methoxy-N-methyl-succinamic acid tert-butylester (10.00 g, 27.29 mmol) in Ethyl ether (200 mL) at −5° C. was addeddropwise 0.5 equiv of 1.0M Lithium aluminum hydride (LAH) in ethyl ether(13.65 mL, 13.65 mmol). After stirring at −5° C. to room temperature for1 hr, the reaction mixture was treated with 5% KHSO₄ (200 mL), stirredfor 5 min and then partitioned between EtOAc/water. The organic phasewas washed with saturated NaCl solution (200 mL), dried over Na₂SO₄ andevaporated to dryness. The residue was purified by flask columnchromatography on silica gel eluting with EtOAc/Hexane (20-80%) to givethe tittle compound (6.00 g, 71%) as a clear oil. TLC (40% EtOAc/Hexane)R_(f)=0.38; ¹HNMR (CDCl₃) δ 9.62 (s, 1H), 7.39 (s, 5H), 5.88 (d,J=7.8Hz, 1H), 5.15 (s, 2H), 4.43-4.37 (dt, J=9.3, 4.5Hz, 1H), 3.01-2.94(dd, J=17.4, 4.8Hz, 1H), 2.80-2.73 (dd, J=17.4, 4.8Hz, 1H), 1.42 (s,9H). MS(ES) for C₁₆H₂₁NO₅ (MW=307.14): positive 308 (MH⁺).

[0423] Part J: 3-Benzyloxycarbonylamino-4,4-diethoxy-butyric acidtert-butyl ester

[0424] To a suspension of 3-Benzyloxycarbonylamino-4-oxo-butyric acidtert-butyl ester (4.46 g, 14.53 mmol) in Ethanol (40 mL) at 0° C. wasadded 0.20 equiv TsOH.H₂O (0.55 g, 2.91 mmol) and followed by dropwise8.0 equiv of CH(OEt)₃ (19.34 mL, 116.25 mmol). After stirring at 0° C.to room temperature for 16 hrs, the reaction mixture was treated withsaturated NaHCO₃ (100 mL), stirred for 5 min and then partitionedbetween EtOAc/water. The organic phase was washed with saturated NaClsolution (200 mL), dried over Na₂SO₄ and evaporated to dryness. Theresidue was purified by flask column chromatography on silica geleluting with EtOAc/Hexane (20%) to give the tittle compound (4.88 g,88%) as a clear oil. TLC (30% EtOAc/Hexane) R_(f)=0.33; ¹HNMR (CDCl₃) δ7.38 (m, 5H), 5.28 (d, J=9.9 Hz, 1H), 5.15-5.05 (dd, J=18.0, 12.3 Hz,2H), 4.48 (d, J=3.6 Hz, 1H), 4.21-4.16 (m, 1H), 3.75-3.65 (m, 2H),3.59-3.47 (m, 2H), 2.59-2.46 (dd, J=15.6, 5.7 Hz, 2H), 1.42 (s, 9H),1.21-1.16 (t, J=6.9 Hz, 3H).

[0425] Part K: 3-Amino-4,4-diethoxy-butyric acid tert-butyl ester

[0426] To a suspension of 3-Benzyloxycarbonylamino-4,4-diethoxy-butyricacid tert-butyl ester (0.50 g, 1.31 mmol) in Ethyl acetate (50 mL) atroom temperature was added 10 mol% Pd/C (˜0.05 g). The reaction flaskwas purged between H₂ (1 atm) and vacuum three times before stirringunder H₂ (I atm) at room temperature. After stirring at room temperaturefor 3 hrs, the reaction mixture was filtered through celite and thenevaporated to dryness to give the title compound (0.32 g, 98%) as aclear oil. ¹HNMR (CDCl₃) δ 4.29 (d, J=5.4 Hz, 1H), 3.79-3.67 (m, 2H),3.61-3.49 (m, 2H), 3.29-3.21 (m, 1H), 2.60-2.53 (dd, J=16.2, 4.2 Hz,1H), 2.30-2.21 (dd, J=16.2, 9.0 Hz, 1H), 1.46 (s, 9H), 1.25-1.19 (m,3H).

[0427] Part L:(3S)-3-{(2S)-2-[(5-Acetylamino-2-tert-butyl-phenylaminooxalyl)-amino]-propionylamino-4,4-diethoxy-butyricacid tert-butyl ester

[0428] To a solution of(2S)-2-[(5-Acetylamino-2-tert-butyl-phenylaminooxalyl)-amino]-propanepanoicacid (0.16 g, 0.46 mmol) in CH₂Cl₂/NMP (1:1, 3 mL) at room temperatureunder N₂ was added 1.5 equiv HATU (0.26 g, 0.69 mmol). The mixture wasstirred at room temperature under N₂ for 45 min before addition of3-Amino-4,4-diethoxy-butyric acid tert-butyl ester (0.13 g, 0.51 mmol),followed by Et₃N (0.19 mL, 1.38 mmol). After stirring at roomtemperature for 3hrs, the mixture was partitioned between EtOAc-water.The organic phase was washed with water, 5% KHSO₄, saturated NaHCO₃, andsaturated NaCl solutions, dried over anhydrous Na₂SO₄ and evaporated togive crude title compound. The residue was purified by flask columnchromatography on silica gel eluting with EtOAc/Hexane (30-70%) to givethe title compound (0.26 g, 99%) as a white foam. TLC (70% EtOAc/Hexane)R_(f)=0.31; ¹HNMR (CDCl₃) δ 9.80 (bs, 1H), 8.81 (d, J=8.1 Hz, 1H), 8.54(s, 1H), 8.00 (d, J=2.1 Hz, 1H), 7.86-7.83 (dd, J=8.7, 2.1 Hz, 1H),7.37-7.34 (d, J=8.7 Hz, 1H), 6.85 (d, J=8.7 Hz, 1H), 4.61-4.33 (m, 3H),3.77-3.39 (m, 4H), 2.50-2.46 (dd, J=6.3, 2.4 Hz, 2H), 2.18 (s, 3H). 1.50(d, J=6.9 Hz, 3H), 1.46 (s, 9H); MS(ES) for C₂₉H₄₆N₄O₈ (MW=578.70):positive 596 ([M+NH₄]⁺).

[0429] Part M:N-(5-Acetylamino-2-tert-butyl-phenyl)-N′-[1-(2-hydroxy-5-oxo-tetrahydrofuran-3-ylcarbamoyl)-ethyl]-oxalamide

[0430] The starting material,(3S)-3-f(2S)-2-[(5-Acetylamino-2-tert-butyl-phenylaminooxalyl)-amino]-propionylamino}-4,4-diethoxy-butyricacid tert-butyl ester (0.24 g, 0.42 mmol) was stirred inWater/TFA/CH₂Cl₂ (1:1:2, 4 mL) at room temperature. After stirring for2hrs, the reaction solution was concentrated in vacuo to drynessyielding the crude title compound. The residue was purified bypreparative HPLC (C,₈ column eluding with 10-90% of 0.1% aqueous formicacid/ACN over 60 min) to give the title compound (0.09g, 46%) as a whitesolid. MS(ES) for C₂₁H₂₈N₄O₇ (MW=448.47): positive 449 ([MH⁺]).

EXAMLE 221

[0431]

[0432]N-(5-Acetylamino-2-Tert-Butyl-Phenyl)-N′-[1-(2-Ethoxy-5-Oxo-Tetrahydro-Furan-3-Ylcarbamol)-Ethyl]-Oxalamide

[0433] Part A: (2-Ethoxy-5-oxo-tetrahydro-furan-3-yl)-carbamic acidbenzyl ester

[0434] To a suspension of 3-Benzyloxycarbonylamino-4,4-diethoxy-butyricacid tert-butyl ester (4.61 g, 12.09 mmol) in 20% anhydrous TFA/CH₂Cl₂(20 mL) at room temperature was added Anisole (0.4 mL). After stirringat room temperature for 1 hr. the reaction mixture was evaporated todryness to give the crude title compounds as a clear oil. The residuewas purified by flask column chromatography on silica gel eluting withEtOAc/Hexane (10-30%) to give:

[0435] (2S, 3 S)-(2-Ethoxy-5-oxo-tetrahydro-furan-3-yl)-carbamic acidbenzyl ester (1.54g, 46%) as a clear oil. TLC (30% EtOAc/Hexane)R_(f)=0.52; ¹HNMR (CDCl₃) δ 7.38 (m, 5H), 5.40 (br.s, 1H), 5.11 (br.s,2H), 5.00 (br.s, 2H), 4.21 (t, J=6.6 Hz, 1H), 3.90-3.80 (dd, J=7.2, 2.4Hz, 1H), 3.66-3.61 (d, J=8.1 Hz, 1H), 2.90-2.81 (dd, J=17.1, 8.4 Hz,1H), 2.51-2.41 (dd, J=17.4, 10.5 Hz, 1HI, 1.24 (t, J=6.9 Hz, 3H). MS(ES) for C₁₄H₁₇NO₅ (MW=279.29): positive 278 ([MH⁺]).

[0436] (2R, 3 S)-(2-Ethoxy-5-oxo-tetrahydro-furan-3-yl)-carbamic acidbenzyl ester (0.89g, 26%) as a clear oil. TLC (30% EtOAc/Hexane)R_(f)=0.42 ¹HNMR (CDCl₃) δ 7.37 (m, 5H), 5.43 (d, J=5.1 Hz, 1H), 5.11(br.s, 2H), 5.34 (d, J=9.3 Hz, 1H), 4.61-4.50 (m, 1H), 3.95-3.85 (dq,J=9.3, 6.9 Hz, 1H), 3.68-3.58 (dq, J=9.3, 6.9 Hz, 1H), 2.90-2.81 (dd,J=17.1, 8.4 Hz, 1H), 2.51-2.41 (dd, J=17.4, 10.5 Hz, 1H), 1.24 (t, J=6.9Hz, 3H). MS(ES) for C₁₄H₁₇NO₅ (MW=279.29): positive 278 ([MH⁺]).

[0437] Part B: (4S. 5S)-4-Amino-5-ethoxy-dihydro-furan-2-one

[0438] To a suspension of (2S,3S)-(2-Ethoxy-5-oxo-tetrahydro-furan-3-yl)-carbamic acid benzyl ester(0.43 g, 1.54 mmol) in Ethyl acetate (50 mL) at room temperature wasadded 10 mol% Pd/C (˜0.05 g). The reaction flask was purged between H₂(1 atm) and vacuum three times before stirring under H₂ (1 atm) at roomtemperature. After stirring at room temperature for 3 hrs, the reactionmixture was filtered through celite and then evaporated to dryness togive the title compound (0.21 g, 94%) as a clear oil. ¹HNMR (CDCl₃) δ5.17 (d, J=1.2 Hz, 1H), 3.93-3.82 (dq, J=9.6, 7.2 Hz, 1H), 3.67-3.57(dq, J=9.3, 6.9 Hz, 1H), 2.95-287 (dd, J=17.7, 6.9 Hz, 1H), 2.56-2.19(dd, J=17.4, 2.7 Hz, 1H), 1.26-1.21 (t, J=7.2 Hz, 3H). MS(ES) forC₆H₁₁NO₃ M=145.16): positive 146 ([MH⁺]).

[0439] Part C: (4S. 5R)-4-Amino-5-ethoxy-dihydro-furan-2-one

[0440] To a suspension of (2R, 3S)-(2-Ethoxy-5-oxo-tetrahydro-furan-3-yl)-carbamic acid benzyl ester(0.22 g, 0.86 mmol) in Ethyl acetate (25 mL) at room temperature wasadded 10 mol% Pd/C (˜0.03 g). The reaction flask was purged between H₂(1 atm) and vacuum three times before stirring under H₂ (1 atm) at roomtemperature. After stirring at room temperature for 3 hrs, the reactionmixture was filtered through celite and then evaporated to dryness togive the title compound (0.12 g, 97%) as a clear oil. ¹HNMR (CDCl₃) δ5.31 (d, J=5.1 Hz, 1H), 3.98-3.87 (dq, J=9.6, 7.2 Hz, 1H), 3.76-3.59 (m,2H), 3.52-3.45 (q, J=7.2 Hz, 1H), 2.72-2.64 (dd, J=17.4, 8.1 Hz, 1H),2.45-2.36 (dd, J=17.1, 10.2Hz, 1H), 1.29-1.24 (t, J=6.9 Hz, 3H). MS(ES)for C₆H₁₁NO₃ (MW=145.16): positive 146 ([MH⁺]).

[0441] Part D:N-(5-Acetylamino-2-tert-butyl-phenyl)-N′-[1-(2-ethoxy-5-oxo-tetrahydro-furan-3-ylcarbamoyl)ethyl]-oxalamide

[0442] To a solution of(2S)-2-[(5-Acetylamino-2-tert-butyl-phenylaminooxalyl)-amino]-propanoicacid (0.24 g, 0.68 mmol) in CH₂Cl₂/NMP (1:1, 3 mL) at room temperatureunder N₂ was added 1.5 equivO-benzotriazol-1-yl-N,N,N′,N′-tetramethyluronium hexafluorophosphate(HBTU) (0.26 g, 0.69 mmol). The mixture was stirred at room temperatureunder N₂ for 45 min before added (4S,5S)-4-Amino-5-ethoxy-dihydro-furan-2-one (0.12 g, 0.82 mmol) andfollowed by Et₃N (0.24 mL, 1.70 mmol). After stirring at roomtemperature for 16 hrs, the mixture was partitioned between EtOAc-water.

[0443] The organic phase was washed with water, and saturated NaClsolutions, dried over anhydrous Na₂SO₄ and evaporated to give crudetitle compound. The residue was purified by flask column chromatographyon silica gel eluding with EtOAc/Hexane (60-80%) to give the titlecompound (0.14 g, 43%) as a white solid. TLC (100% EtOAc) R_(f)=0.51;¹HNMR (CDCl₃) δ 9.82 (t, J=10.5 Hz, 1H), 8.85-8.44 (m, 2H), 8.00-7.94(m, 1H), 7.80-7.68 (m, H1H), 7.38-7.34 (m, 1H), 5.41-5.33 (dd, J=22.2,4.8 Hz, 1H), 4.74-4.63 (m, 1H), 4.59-4.35 (m, 1H), 3.88-3.77 (dq, J=9.6,7.2 Hz, 1H), 3.70-3.56 (m, 1H), 3.39-3.22 (m, 1H). 3.03-2.88 (ddd,J=18.3, 7.8, 2.4 Hz, 1H), 2.56-2.35 (m, 1H), v 2.19 (s, 3H), 1.57-1.52(dd, J=7.2, 3.0 Hz, 3H), 1.45 (s, 91); MS(ES) for C₂₃H₃₂N₄O₇(MW=476.52): positive 477 ([MH⁺]).

EXAMPLE 222

[0444]

[0445]N-(5-Acetylamino-2-Tert-Butyl-Phenyl)-N′-[1-(2-Ethoxy-5-Oxo-Tetrahydro-Furan-3-Ylcarbamoyl)-Ethyl]-Oxalamide

[0446] To a solution of(2S)-2-[(5-Acetylamino-2-tert-butyl-phenylaminooxalyl)-amino]-propanoicacid (0.08 g, 0.23 mmol) in CH₂Cl₂/NMP (1:1, 1 5 mL) at room temperatureunder N₂ was added 1.5 equiv HBTU (0.13 g, 0.35 mmol). Mixture solutionwas stirred at room temperature under N₂ for 45 min before added (4S,5R)-4-Amino-5-ethoxy-dihydro-fuiran-2-one (0.05 g, 0.35 mmol) andfollowed by Et₃N (0.10 mL, 0.69 mmol). After stirring at roomtemperature for 18hrs, the mixture was partitioned between EtOAc-water.The organic phase was washed with water, and saturated NaCl solutions,dried over anhydrous Na₂SO₄ and evaporated to give crude title compound.The residue was purified by flask column chromatography on silica geleluting with EtOAc/Hexane (60-90%) to give the title compound (0.10 g,91%) as a white solid. TLC (100% EtOAc) R_(f)=0.52; ¹HNMR (CDCl₃) δ 9.77(s, 1H), 8.70-8.65 (t, J=8.1 Hz, 1H), 8.48 (s, 1H), 8.00 (s, 1H), 7.82(d, J=11.4 Hz, 1H), 7.39-7.36 (d, J=8.7 Hz, 1H), 7.06 (d, J=7.5 Hz, 1H),1H), 4.49 (d, J=5.1 Hz, 1H), 4.78-4.57 (m, 2H), 3.98-3.86 (m, 1H),3.72-3.57 (m, 1H), 2.49-2.35 (m, 1H), 2.17 (s, 3H), 2.08-1.97 (m, 1H),1.55 (d, J=7.2 Hz, 3H) 1.46 (s, 9H), 1.28-1.22 (t, J=14.1 Hz, 3H).MS(ES) for C₂₃H₃₂N₄O₇ (MW=476.52): positive 477 ([MH⁺]), 499 ([M+Na]⁺).

EXAMPLE 223

[0447]

[0448]N-(2-Tert-Butyl-Phenyl)-N′-[1-(2-Ethoxy-5-Oxo-Tetraydro-Furan-3-Ylcarbamoyl)-Ethyl]-Oxalamide

[0449] Part A: Methyl 2-(2-tert-Butylphenylamino)-2-oxoacetate

[0450] A 3-L, 3-necked round-bottomed flask, equipped with a mechanicstirrer and a thermal probe (under nitrogen) was charged with2-tert-Butylaniline (109 g, 114 mL, 732 mmoles), triethylamine (81.4 g,112 mL, 804 mmoles, 1.1 equiv.) and toluene (600 mL). The resultingmixture was stirred at a moderate speed at −30° C. An additon funnel wascharged with Methyl chlorooxoacetate (100 g, 816 mmoles, 1.11 equiv.)with toluene (200 mL), and the mixture added to the reaction mixture atsuch a rate that the internal batch temperature is less than −20° C.After addition, the reaction was warmed to room temperature for an hour,quenched with water, then partitioned between E,tOAc/water. The aqueousphase was extracted with EtOAc (200 mL), and the combined organic layerswere washed successively with KHSO₄ (200 mL), NaHCO₃ (sat'd) (200 mL)and brine (200 mL), then dried the organic over MgSO₄. The organic phasewas concentrated by rotary evaporation, yielding the title compound as apale yellow solid [Methyl 2-(2-tert-Butylphenylamino)-2-oxoacetate, 160g, 93.1%]. MP 61.7-63.6° C. IR (KBr) 3409, 2954, 1736, 1724, 1530, 1299,1166, 766 cm⁻¹; ¹H NMR (300 MHz, CDCl₃): δ (ppm): 9.20 (br, 1H), 7.97(dd, J=7.8 Hz, J′=1.8 Hz, 1H), 7.43 (dd, J=7.8 Hz, J′=1.8 Hz, 1H),7.31-7.16 (m, 2H), 4.00 (s, 3H), 1.47(s, 9H). The product thus obtainedcan be used directly for the subsequent reaction.

[0451] Part B: 2-(2-tert-Butyl phenylamino)-2-oxoacetic acid

[0452] A 3-L, 3-necked round-bottomed flask, equipped with a mechanicstirrer and a thermal probe (under nitrogen) was charged with Methyl2-(2-tert-Butylphenylamino)-2-oxoacetate (154 g, 655 mmoles) and MeOH(1000 mL). The resulting mixture was stirred at a moderate speed at roomtemperature. A solution of 1N NaOH/MeOH (800 mL) was added dropwise tothe reaction mixture via addition funnel. After 1 hour, agitation wasstopped, and the suspension transferred to a filtration funnel andfiltered to afford a white solid. This solid was then taken up in water(1200 mL) and the pH adjusted to 1.5-2 with addition of conc. HCl withstirring. After mixing for 1 hour, filtration and drying yielded thetitle compound as a white crystalline mass (127 g, 574 mmoles, 87.6%).TLC (silica F254, 3/1 v/v dichloromethane/MeOH, detection 254 nm) showedone spot and HPLC confirmed that. MP 110.9-112.6° C.; IR (KBr) 3405,2973, 1688, 1548, 1300, 757 cm⁻¹; ¹H NMR (300 MHz, CDCl₃): δ (ppm): 9.37(br, 1H), 7.91 (dd, J=7.8 Hz, J′=1.8 Hz, 1H), 7.44 (dd, J=7.8 Hz, J′=1.8Hz, 1H), 7.25 (m, 2H), 1.46 (s, 9H); ¹³C (300 MHz, CDCl₃): δ (ppm):160.87, 155.24, 141.18, 133.19, 126.92, 126.69, 126.63, 124.24, 34.21,30.49.

[0453] Part C: Methyl(2S)-2-({N-[2-(tert-butyl)phenyl]carbamoyl}carbonylamino)propanoate

[0454] A 3-L, 3-necked round-bottomed flask, equipped with a mechanicstirrer and a thermal probe (under nitrogen) was charged with thefollowing solids: 2-(2-tert-Butylphenylamino)-2-oxoacetate (70.008 g,316.402 mmol), alanine methyl ester hydrochloride (44.368 g, 317.862mmol), hydroxybenzotriazole hydrate (HOBT) (47.324 g, 350.210 mmol), and1-[3-(Dimethylamino)propyl]-3-ethylcarbodiimide hydrochloride (EDAC•HCl)(121.668 g, 635.4 mmol). N,N′-dimethylformamide (DMF) (1250 niL) wasadded via graduated cylinder, and the reaction mixture stirred topartially dissolve and suspend the solid reagents. N-methylmorpholine(108 mL, 99.36 g, 982 mmol) was then added in one portion via graduatedcylinder. After stirring at room temperature for 18 hours, the reactionmixture was partitioned between EtOAc (ca. 600 mL) and 1N HCl (ca. 500mL). The aqueous phase was washed two times with EtOAc (ca. 300 mL), andthe combined organic layers were washed successively with 1N HCl (ca.300 mL), H₂O (ca. 300 mL), saturated NaHCO₃ (ca. 300 mL), and brine (ca.300 mL). The resulting pale yellow organic solution was dried overMgSO4, then concentrated to yield a pale yellow crystalline mass (95 g,97%). TLC: (silica, 1:1 hexanes/EtOAc) R_(f): 0.54. MP: 60.1-62.1° C.;IR (KBr) 3284, 1747, 1662, 1503, 1216, 755 cm⁻¹; ¹H NMR: (CDCl₃, TMS) δ9.55 (br, 1H), 8.01 (dd, J=7.8 Hz, J′=1.8 Hz, 1H), 7.42 (dd, J=7.8 Hz,J′=2.6 Hz, 1H), 7.15-7.31 (m, 2H), 4.64 (p, J=7.5 Hz, 1H), 3.80 (s, 3H),1.54 (d, J=7.5 Hz, 3H), 1.46 (s, 9H) ppm.

[0455] Part D:(2S)-2-({N-[2-(tert-butyl)phenyl]carbamoyl}carbonylamino)propanoic acid

[0456] A 3-L, 3-necked round-bottomed flask, equipped with a mechanicstirrer and a thermal probe (under nitrogen) was charged with methyl(2S)-2-({N-[2-(tert-butyl)phenyl]-carbamoyl}carbonylamino)propanoate(85.0 g, 276.5 mmol), dissolved in dioxane (664 mL) and the resultingsolution cooled in an ice water bath to 8° C. as monitored by aninternal temperature probe. To this was added 1N LiOH (332 mL) in threeapproximately equal portions such that the temperature did not exceed10° C. with stirring. The bath was removed and stirring continued over2.5 hours (as the reaction comes to room temperature) at which point TLCanalysis (silica gel, 1:1 EtOAc/hexanes) of the reaction mixture showedthat the starting material had been consumed. The reaction mixture waspartitioned between IN HCl (300 mL) and EtOAc (500 mL) in a 2Lseparatory funnel and separated. The aqueous phase tested to pH 1-2 bytest strip. The resulting aqueous fraction was washed with three (3)additional portions of EtOAc (3×200 mL), then the organic fractions werecombined and washed consecutively with 10% citric acid solution (200 mL)and brine (200 mL). The organic phase was then concentrated to a mobilegold oil and further dried under high vacuum 20 h to yield a crudeyellow solid mass (about 90 g, very moist). The solid mass wasrecrystallized from EtOAc/Hexanes, and dried to afford a highlycrystalline white solid (58.545 g, 200 mmol, 72.4%). TLC: (silica, 19:1CH₂Cl₂/MeOH) R_(f): 0.18; MP: 164.4-166.1° C.; IR (KBr) 3323, 1719,1677, 1517, 1448, 1248, 762 cm⁻¹; ¹H NMR: (300 MHz, CDCl₃) δ 9.57 (br,1H), 8.10 (d, J=7.8 Hz, 1H), 7.97 (dd, J=7.8 Hz, J′=1.8 Hz, 1H), 7.43(dd, J=7.8 Hz, J′=1.8 Hz, 1H), 7.25-7.30 (m, 1H), 7.15-7.21 (m, 1H),4.67 (p, J=7.5 Hz, 1H), 1.60 (d, J=7.2 Hz, 3H), 1.46 (s, 9H) ppm; ¹³CNMR: (300 MHz, CDCl₃) δ 175.71, 159.97, 156.33, 140.92, 133.77, 126.79,126.47, 126.01, 124.01, 48.31, 34.21, 30.42, 17.50; Elemental Analysis:Calcd for C₁₅H₂₀N₂O₄: C, 61.63; H, 6.90; N, 9.58. Found: C, 61.80; H,6.95; N, 9.50.

[0457] Part E: (3S)-3-1(2S)-2-[(2-tert-Butyl-phenylaminooxalyl)-amino]-propionylamino}-4,4-diethoxy-butyricacid tert-butyl ester

[0458] To a solution of(2S)-2-[(2-tert-butyl-phenylaminooxalyl)-amino]-propanoic acid (1.20 g,4.10 mmol) in CH₂Cl₂/NMP (1:1, 10 mL) at room temperature under N₂ wasadded 1.5 equiv HATU (0.26 g, 6.15 mmol). The mixture solution wasstirred at room temperature under N₂ for 45 min before addition of3-Amino-4,4-dielhoxy-butyric acid tert-butyl ester (1.01 g, 4.10 mmol),followed by Et₃N (1.71 mL, 12.30 mmol). After stirring at roomtemperature for 16hrs, the mixture was partitioned between EtOAc-water.The organic phase was washed with water, 5% KHSO₄, saturated NaHCO₃, andsaturated NaCl solutions, dried over anhydrous Na₂SO₄ and evaporated togive crude title compound. The residue was purified by flask columnchromatography on silica gel eluding with EtOAc/Hexane (20%) to give thetittle compound (1.93 g, 90%) as a white foam. TLC (40% EtOAc/Hexane)R_(f)=0.60; 1HNMR (CDCl₃) δ 9.54 (s, 1H), 8.09 (d, J=7.5 Hz, 1H), 8.00(dd, J=8.1, 1.5 Hz, 1H), 7.42 (dd, J=9.6, 1.5 Hz, 1H), 7.27 (dt, J:=7.2,0.9 Hz, 1H), 7.17 (dd, J=7.8, 1.5 Hz, 1H), 6.52 (d, J=9.0 Hz, 1H), 4.53(d, J=3.9, 1H), 4.50-4.37 (m, 2H), 3.77-3.64 (m, 2H), 3.60-3.46 (m, 2H).2.52 (d, J=6.9 Hz, 2H), 1.51-1.42 (m, 12H), 1.29-1.16 (m, 6H); MS(ES)for C₂₇H₄₃N₃O₇ (MW=521.66): negative 521([M−H]⁻).

[0459]

[0460] Part F:(2-tert-Butyl-phenyl)-N′-[1-(2-hydroxy-5-oxo-tetrahydro-furan-3-ylcarbamoyl)-ethyl]-oxalamide

[0461] (3S)-3-{(2S)-2-[(2-tert-Butyl-phenylaminooxalyl)-amino]-propionylamino}-4,4-diethoxy-butyricacid tert-butyl ester (1.39 g, 2.66 mmol) was stirred inWater/TFA/CH₂Cl₂ (30 mL, 1:1:2) at room temperature. After stirring for2 hrs, the reaction solution was concentrated in vacuo until dryness togive crude title compound. The residue was purified by preparative HPLC(C₁₈ column eluding with 30-90% of 0.1% aqueous formic acid/ACN over 60min) to give the title compound (0.91 g, 87%) as a white solid. LCMS forC₁₉H₂₅N₃O₆ (MW=391.42): negative 390.32 ([MH⁻]). Purity assay usingreverse phase Zorbax C8 4.6×150 mm Rx column over 30 min., withretention time t=10.75 min.

EXAMPLE 225

[0462]

[0463]N-(2-Tert-Butyl-Phenyl)-N′-[1-(2-Ethoxy-5-Oxo-Tetrahydro-Furan-3-Ylcarbamoyl)-Ethyl]-Oxalamide

[0464] To a solution of(2S)-2-[(2-terf-Butyl-phenylaminooxalyl)-amino]-propanoic acid (1.00 g,mmol) in CH₂Cl₂/NMP (1:1, 3 mL) at room temperature under N₂ was added1.5 equiv HBTU (0.26 g, 3.42 mmol). Mixture solution was stirred at roomtemperature under N₂ for 45 min before added (4S,5S)-4-Amino-5-ethoxy-dihydro-furan-2-one (0.42 g, 2.86 mmol) andfollowed by Et₃N (1.20 mL, 8.58 mmol). After stirring at roomtemperature for 16 hrs, the mixture was partitioned between EtOAc-water.The organic phase was washed with water, then brine, dried overanhydrous Na₂SO₄ and evaporated to give crude title compound. Theresidue was purified by flask column chromatography on silica geleluting with EtOAc/Hexane (10-40%) to give the title compound (0.53 g,44%) as a white solid. TLC (50% EtOAc/Hexane) Rf=0.45; ¹HNMR (CDCl₃) δ9.52 (s, 1H), 7.95 (m, 2H), 7.43 (dd, J=7.8, 1.5 Hz, 1H), 7.31-7.25 (dt,J=7.2, 1.2 Hz, 1H), 7.22-7.16 (dt, J=7.5, 1.5Hz, 1H), 6.88 (d, J=6.9 Hz,1H), 5.37 (s, 1H), 4.50-4.39 (m, 2H), 3.89-3.79 (dq, J=9.6, 7.2 Hz, 1H),3.69-3.59 (dq, J=9.3, 6.6 Hz, 1H), 3.07-2.98 (dd, J=18.0, 7.5 Hz, 1H).2.39-2.33 (dd, J=19.5, 1.2 Hz, 1H), 1.50 (d, J=7.2 Hz, 1H), 1.46 (s,9H), 1.23 (t, J=7.2 Hz, 3H); LCMS for C₂₁H₂₉N₃O₆ (MW=419.47): negative418 (MH⁻), positive 420 ([MH⁺]).

EXAMPLES 226-312

[0465] Starting with (3S)-3-[N-(9-fluorenylmethoxycarbonyl)alanyl]amino-4-oxobutanoic acid(tert-butyl) ester semicarbazonyl-4[2′-(4-ethyl-phenoxyacetyl)]aminomethylpolystene (see Example 204, Part A) and following the methodsdescribed in Example 204, Part B, or by the procedures set forth inExamples 220-225, the compounds shown below in Table 12 were prepared:TABLE 12

MS(ES) Ex. R¹ Formula MW Pos neg 226 (4-Ac)Ph C₁₇H₁₉N₃O₇ 377.35 — 376(M− H) 227 (4-OH)Ph C₁₅H₁₇N₃O₇ 351.32 — — 228 (3,5-di-Cl-4-OH)PhC₁₅H₁₅Cl₂N₃O₇ 420.21 — 418/420(M − H) 229 (3-CF3)PhCH2 C₁₇H₁₈F₃N₃O₆417.34 — — 230 (2-F)PhCH2 C₁₆H₁₈FN₃O₆ 367.33 — — 231 (2,4-di-Cl)PhC₁₇H₁₉Cl₂N₃O₆ 432.26 — — 232 (3,4-di-OCH3)Ph C₁₈H₂₃N₃O₈ 409.40 — — 233CH3 C₁₀H₁₅N₃O₆ 273.25 — — 234 S-(1-naphthyl)CH(CH3) C₂₁H₂₃N₃O₆ 413.43 —— 235 R-(1-naphthyl)CH(CH3) C₂₁H₂₃N₃O₆ 413.43 — — 236 (2-(1-naphthyl))PhC₂₅H₂₃N₃O₆ 461.47 484(M + Na) 460(M − H) 237 5-Ph-3-pyrazolyl C₁₈H₁₉N₅O₆401.38 — 400(M − H) 238 5,6,7,8-tetrahydro-1-naphthyl C₁₉H₂₃N₃O₆ 389.41— — 239 Ph2CH C₂₂H₂₃N₃O₆ 425.44 — — 240 (2-I)Ph C₁₅H₁₆IN₃O₆ 461.21 — —241 (2,3,5,6-tetra-Cl)Ph C₁₅H₁₃Cl₄N₃O₆ 473.10 — — 242 (4-Ph)PhC₂₁H₂₁N₃O₆ 411.41 — — 243 (2-PhO)Ph C₂₁H₂₁N₃O₇ 427.41 — — 244 2-naphthylC₁₉H₁₉N₃O₆ 385.38 — — 245 1,2,3,4-tetrahydro-1-naphthyl C₁₉H₂₃N₃O₆389.41 — — 246 1-naphthylCH2 C₂₀H₂₁N₃O₆ 399.40 — — 247 1-adamantylC₁₉H₂₇N₃O₆ 393.44 — — 248 4-pyridyl C₁₄H₁₆N₄O₆ 336.30 — — 249(2,3,4,5,6-penta-F)Ph C₁₅H₁₂F₅N₃O₆ 425.27 — — 250 (2-F-4-I)PhC₁₅H₁₅FIN₃O₆ 479.20 — — 251 1,1,3,3-tetramethylbutyl C₁₇H₂₉N₃O₆ 371.43 —— 252 Ph(CH2)2 C₁₇H₂₁N₃O₆ 363.37 — — 253 n-heptyl C₁₆H₂₇N₃O₆ 357.41 — —254 (4-n-heptyl)Ph C₂₂H₃₁N₃O₆ 433.50 — — 255 (2,5-di-tBu)Ph C₂₃H₃₃N₃O₆447.53 — — 256 (2-PhCH2)Ph C₂₂H₂₃N₃O₆ 425.44 — — 257(2-pyrrolidin-1yl-5-CF3)Ph C₂₀H₂₃F₃N₄O₆ 472.42 — — 2582,3,5,6-tetra-F-4-pyridyl C₁₄H₁₂F₄N₄O₆ 408.27 — — 259 (2-Ph)PhC₂₁H₂₁N₃O₆ 411.41 450(M + K) 410(M − H) 260 (3,4,5-tri-Cl)PhC₁₅H₁₄Cl₃N₃O₆ 438.65 — — 261 (4-OCH3)Ph C₁₆H₁₉N₃O₇ 365.34 — — 262PhNH(CS)NH C₁₆H₁₈N₄O₆S 394.40 — — 263 (2,4-di-Br)Ph C₁₅H₁₅Br₂N₃O₆ 493.11— — 264 6-quinolinyl C₁₈H₁₈N₄O₆ 386.36 — — 265 (3,4,5-tri-OCH3)PhCH2C₁₉H₂₅N₃O₉ 439.42 — — 266 (4-CH3)Ph C₁₆H₁₉N₃O₆ 349.34 372(M + Na) 348(M− H) 267 (2-F)Ph C₁₅H₁₆FN₃O₆ 353.31 — — 268 (2-Br-4-Cl-6-F)PhC₁₅H₁₄BrClFN₃O₆ 466.65 — — 269 PhCH2 C₁₆H₁₉N₃O₆ 349.34 — 348(M − H) 270Ph C₁₅H₁₇N₃O₆ 335.32 — — 271 (2,6-di-F)Ph C₁₅H₁₅F₂N₃O₆ 371.30 — — 272(2,3,4,6-tetra-F)Ph C₁₅H₁₃F₄N₃O₆ 407.28 — — 273 (2,4-di-Cl)PhC₁₅H₁₅Cl₂N₃O₆ 404.21 — 402/404(M − H) 274 (2-CF3)Ph C₁₆H₁₆F₃N₃O₆ 403.31426(M + Na) 402(M − H) 275 5-indanyl C₁₈H₂₁N₃O₆ 375.38 — — 276 (3-OPh)PhC₂₁H₂₁N₃O₇ 427.41 — — 277 4-Cl-1-naphthyl C₁₉H₁₈ClN₃O₆ 419.82458/460(M + K) 418/420(M − H) 278 1,4-benzodioxan-6-yl C₁₇H₁₉N₃O₈ 393.35— — 279 (2-Cl)Ph C₁₅H₁₆ClN₃O₆ 369.76 — 368/370(M − H) 280 (2-Br)PhC₁₅H₁₆BrN₃O₆ 414.21 414/416(M + H), 412/414(M − H), 434/436(M + Na),526/628(M + TFA) 452/454(M + K) 281 1-naphthyl C₁₉H₁₉N₃O₆ 385.38 — — 282(2-tBu)Ph C₁₉H₂₅N₃O₆ 391.42 — 390(M − H) 283 (2-tBu-5-NHAc)Ph C₂₁H₂₈N₄O₇448.48 449(M + H) 284 5-isoquinolinyl C₁₈H₁₈N₄O₆ 386.36 387(M + H) 385(M− H) 285 1-indanyl C₁₈H₂₁N₃O₆ 375.38 — — 286 (2-F)Ph(CH2)2 C₁₇H₂₀FN₃O₆381.36 — — 287 (2,4-di-F)PhCH2 C₁₆H₁₇F₂N₃O₆ 385.32 408(M + Na) 385(M −H), 498 (M + TFA) 288 (2,4-di-Cl)PhCH2 C₁₆H₁₇Cl₂N₃O₆ 418.23 — — 289PhNNH C₂₁H₂₂N₄O₆ 426.43 — — 290 PhCONH C₁₆H₁₈N₄O₇ 378.34 — — 291PhCH2(Ph)N C₂₂H₂₄N₄O₆ 440.46 — — 292 Ph(Me)N C₁₆H₂₀N₄O₆ 364.36 — 439(M −H) 293 PhSO2 C₁₅H₁₇N₃O₈S 399.38 — — 294 PhCH2O C₁₆H₁₉N₃O₇ 365.34 — — 295(2,6-di-Cl)Ph C₁₅H₁₆Cl₂N₄O₆ 419.22 — — 296 (4-Me)Ph C₁₆H₂₀N₄O₆ 364.36 —— 297 Ph2NCONH C₂₂H₂₃N₅O₇ 469.45 — — 298 (4-Cl)Ph C₁₅H₁₇ClN₄O₆ 384.78384/386(M + H) 383/385(M − H) 299 (4-NO2)PhCONH C₁₆H₁₇N₅O₉ 423.34 — —300 (2,4-di-NO2)PhNH C₁₅H₁₆N₆O₁₀ 440.33 — — 301 PhNHCONH C₁₆H₁₉N₅O₇393.36 — — 302 (2-NO2)PhCONH C₁₆H₁₇N₅O₉ 423.34 — — 303(2,3,4,5,6-penta-F)PhNH C₁₅H₁₃F₅N₄O₆ 440.28 — — 304 (4-Br)PhNHC₁₅H₁₇BrN₄O₆ 429.23 — — 305 (4-NO2)PHCHN C₁₆H₁₇N₅O₈ 407.34 — 406(M − H)306 PhNH C₁₅H₁₈N₄O₆ 350.33 — 349(M − H) 307 (2,3,5,6-tetra-F)PhNHC₁₅H₁₄F₄N₄O₆ 422.29 — 421(M − H) 308 (4-F)PhNH C₁₅H₁₇FN₄O₆ 368.32 —367(M − H) 309 (3-Cl-4-Me)PhNH C₁₆H₁₉ClN₄O₆ 398.80 397/398/399 397(M −H) (M + H) 310 (3-Cl)PhNH C₁₅H₁₇FN₄O₆ 368.32 — 367(M − H) 3113-[2-ethyl-4(3H)]quinazolinonyl C₁₉H₂₁N₅O₇ 431.40 — — 312 (3-Br)PhCONHC₁₇H₂₁BrN₄O₇ 473.28 471(M − H)

EXAMPLES 313-318

[0466] By the procedures disclosed in Examples 226-312, but startingwith the corresponding tertiary amine, the compounds shown below inTable 13 were also prepared: TABLE 13

MS(ES) Ex. (R¹)(R^(1′))N— Formula MW Pos neg 313 (1-naphthyl)(Me)N—C₂₀H₂₁N₃O₆ 399.40 — — 314 Ph₂N— C₂₁H₂₁N₃O₆ 411.41 — — 315 1-pyrrolidineC₂₀H₂₁F₄N₃O₇ 491.13 — 509(M + NH₄) 429(M +H) 316 1-piperidineC₂₁H₂₃F₄N₃O₇ 505.15 504(M + H) — 317 (1-naphthyl)(Me)N— C₂₀H₂₁N₃O₆399.40 — — 318 Ph₂N— C₂₁H₂₁N₃O₆ 411.41 — —

EXAMPLES 319-394

[0467] By the procedures disclosed in Examples 22, but starting with2-(9H-fluoren-9ylmethoxycarbonylamino)-succinic acid 4-tert-butylesterand the appropriate alcohol, the compounds shown below in Table 14A werealso made: TABLE 14A

Ex. R¹ R^(1′) R² Formula MW MS (ES) 319 5-AcNH-2-tBuPh H ethylC₂₃H₃₂N₄O₇ 476.53 477(M + H), 499(M + Na) 320 2-BrPh H ethylC₁₇H₂₀BrN₃O₆ 422.27 480/482(M + K), 464/466(M + Na), 442/444(M + H) 3212-BnPh H ethyl C₂₄H₂₇N₃O₆ 453.49 476(M + Na), 492(M + K)

[0468] By the above procedures, the compounds listed in Table 14B mayalso be made: TABLE 14A Ex. R¹ R^(1′) R² 322 2-trifluoromethyl-Ph Hmethyl 323 2-BzPh H benzyl 324 2-trifluoromethyl-Ph H ethyl 3251-naphthyl methyl methyl 326 1-naphthyl methyl methyl 3272-trifluoromethyl-Ph H tBu 328 methylPh H H 329 PhPh H methyl 330 PhPh Hethyl 331 3,4-dimethoxyBr H methyl 332 3,4-dimethoxyBr H ethyl 3331-naphthalen-1-yl-ethyl H methyl 334 1-naphthalen-1-yl-ethyl H ethyl 3353,4,5-trimethoxyPh H methyl 336 3,4,5-trimethoxyPh H ethyl 3372-(2-F-Ph)-ethyl H methyl 338 2-(2-F-Ph)-ethyl H ethyl 339 1-naphthyl Hmethyl 340 1-naphthyl H ethyl 341 2-methyl Ph H H 342 2,6-diF-Ph Hmethyl 343 2,6-diF-Ph H ethyl 344 2,6-diF-Ph H tBu 345 2,6-diF-Ph H Bn346 Br H ethyl 347 Br H methyl 348 Br H tBu 349 Br H Bn 350 2,4-diF-Ph HtBu 351 2-PhPh H tBu 352 2-BrPh H tBu 353 1-naphthalen-1-yl-ethyl H tBu354 2-trifluromethylPh H tBu 355 3,4,5-trimethoxy-Ph H tBu 3563-trifluormethylPh H tBu 357 2-(2-F-Ph)-ethyl H tBu 358 1-naphthyl H tBu359 2-I-Ph H methyl 360 2-I-Ph H ethyl 361 2-I-Ph H tBu 362 2-I-Ph H Bn363 2-Br-Ph H methyl 364 2-Br-Ph H ethyl 365 2-Br-Ph H tBu 366 2-Br-Ph HBn 367 1-napthyl methyl Bn 368 3-trifluoromethylPh H Bn 3693-trifluoromethylPh H methyl 370 3-trifluoromethylPh H ethyl 371PhCH(Ph) H methyl 372 PhCH(Ph) H ethyl 373 PhCH(Ph) H Bn 374 Ph Ph Bn375 Ph Ph ethyl 376 Ph Ph methyl 377 2-BrPh H methyl 3782-trifluoromethylPh H Bn 379 2-tBuPh H methyl 380 2-Ph-Ph H Bn 3813,4-dimethoxyPh H Bn 382 1-naphthalin-1-yl-ethyl H Bn 3833,4,5-trimethoxyPh H Bn 384 2-(3-F-Ph)-ethyl H Bn 385 1-naphthyl H Bn386 2,4-diF-Br H Bn 387 2,4-diF-Br H tBu 388 2,4-diF-Br H ethyl 3892,4-diF-Br H methyl 390 Br H tBu 391 3,4-dimethoxy-Br H tBu 3921-naphthyl methyl tBu 393 PhCH(Ph) H tBu 394 Ph Ph tBu

EXAMPLES 395-397

[0469] By the procedurees disclosed in Examples 193-200, but startingwith (N-9-fluorenylmethoxycarbonyl)-tert-butyl glycine, the compoundsshown below in Table 15 were made: TABLE 15

MS(ES) Ex. R¹ = Formula MW Pos neg 395 5-indanyl C₂₈H₂₉F₄N₃O₇ 595.5466 —594(M − H) 396 (2,3,5,6-tetra-Cl)Ph C₂₅H₂₁Cl₄F₄N₃O₇ 693.2624 —690/692/694(M − H) 397 (2-Br)Ph C₂₅H₂₄BrF₄N₃O₇ 634.3781 — 632/634(M − H)

EXAMPLES 398-419

[0470] By the procedures disclosed in Examples 5-21, but starting withthe appropriate amino acid and oxamic acid, the compounds shown in Table16 were also made: TABLE 16

MS(ES) Ex. R¹ R² A B Formula MW pos neg 398 (2-tBu)Ph OtBu valineCH₂O(2,3,5,6- C₂₃H₃₉F₄N₃O₇ 653.27 676(M + Na) 652(M − H) tetra-F—Ph) 399(2-tBu)Ph OCH₃ valine CH₂O(2,3,5,6- C₂₉H₃₃F₄N₃O₇ 611.58 634(M + Na)610(M − H) tetra-F—Ph) 646(M + Cl—) 400 5-quinolin-1- OH valineOPOPh₂CH₂ C₃₅H₃₄F₃N₃O₁₀P 758.2 yl 401 (2-Ph)Ph OH alpha-methyl-CH₂O(2,3,5,6- C₃₅H₂₉F₄N₃O₇ 679.19 702(M + Na) phenylalanine tetra-F—Ph)718(M + JK) 402 (2-tBu)Ph OCH₂Ph alanine CH₂O(2,3,5,6- C₃₃H₃₃F₄N₃O₇659.62 660(M + H) 568(M − H) tetra-F—Ph) 403 (2-Br)Ph CH₂O valine OHC₂₄H₂₂BrF₄N₃O₇ 619.06 618/620(M − H) (2,3,5,6- tetraF—Ph) 404(2,3,4,5-tetra- CH₂O valine OH C₂₄H₁₉F₈N₃O₇ 613.11 612(M − H) F)Ph(2,3,5,6- tetraF—Ph) 405 (2-tBu)Ph OCH₂Ph alanine CH₂O(2,3,5,6-C₃₃H₃₂BrF₄N₃O₇ 737.14 737.88/739.8 735.90/737.90 tetra-F-4-Br)Ph (M + H)(M − H) 406 (2-tBu)Ph OH histidine CH₂O(2,3,5,6- C₂₉H₂₉F₄N₃O₇ 635.2636(M + H) tetra-F—Ph) 407 (2-CF₃)Ph OCH₂Ph alanine CH₂O(2,3,5,6-C₃₀H₂₄F₇N₃O₇ 671.15 672(M + H) 670(M − H) tetra-F—Ph) 408 (2-tBu)Ph OHCys(CH₂Ph) CH₂O(2,3,5,6- C₃₃H₃₃F₄N₃O₈ 675.22 674(M − H) tetra-F—Ph) 409(2-tBu)Ph OH Cys CH₂O(2,3,5,6- C₂₆H₂₇F₄N₄O₇ 585.17 584(M − H)tetra-F—Ph) 410 (2-tBu)Ph OH tryptophan CH₂O(2,3,5,6- C₃₄H₃₂F₄N₄O₇684.22 683(M − H) tetra-F—Ph) 411 (2-tBu)Ph OH lysine CH₂O(2,3,5,6-C₂₉H₃₄F₄N₄O₇ 626.24 625(M − H) tetra-F—Ph) 412 2-(2CH₃O- OCH₂Ph alanineCH₂O(2,3,5,6- C₃₆H₃₁F₄N₃O₈ 709.2 Ph)Ph tetra-F—Ph) 413 piperidin-1-ylOCH₂Ph alanine CH₂O(2,3,5,6- C₂₈H₃₀F₄N₄O₇ 610.21 611(M + H) tetra-F—Ph)414 pyrrolidin-1-yl OCH₂Ph alanine CH₂O(2,3,5,6- C₂₆H₂₈F₄N₄O₇ 596.19597.28(M + H) tetra-F—Ph) 415 Cbz OH valine CH₂O(2,3,5,6- C₂₆H₂₅F₄N₃O₉599.15 622(M + Na) 598(M − H) tetra-F—Ph) 416 5-indanyl OH (t-butyl)CH₂O(2,3,5,6- C₂₈H₂₉F₄N₃O₇ 595.55 — 594(M − H) glycine tetra-F—Ph) 417(2,3,5,6-tetra- OH (t-butyl) CH₂O(2,3,5,6- C₂₅H₂₁Cl₄F₄N₃O₇ 693.26 —690/692/694 Cl)Ph glycine tetra-F—Ph) (M − H) 418 (z-Br)Ph OH (t-butyl)CH₂O(2,3,5,6- C₂₅H₂₄BrF₄N₃O₇ 634.38 — 632/634 glycine tetra-F—Ph) (M −H) 419 2-(2- OCH₂Ph alanine CH₂O(2,3,5,6- C₃₆H₃₁F₄N₃O₈ 709.2 710(M + H)708(M − H) CH₃Oph)Ph tetra-F—Ph)

EXAMLES 420-426

[0471] By the procedures disclosed in Example 126, but starting withintermediates having the desired stereochemistry, the compounds shown inTable 17 were also made: TABLE 17

Ala Asp MS(ES) Ex. R² stereochemistry stereochemistry Formula M/W posneg 420 OBn R R C₃₃H₃₃F₄N₃O₇ 659.62 660(M + H) 658(M − H) 421 OBn R SC₃₃H₃₃F₄N₃O₇ 659.62 660(M + H) 658(M − H) 422 OBn S R C₃₃H₃₃F₄N₃O₇659.62 660(M + H) 658(M − H) 423 H S S C₃₅H₂₉F₄N₃O₇ 569.5 570(M + H)568(M − H) 424 H R R C₃₃H₃₃F₄N₃O₇ 569.5 570(M + H) 568(M − H) 425 H R SC₂₄H₂₂BrF₄N₃O₇ 569.5 570(M + H) 568(M − H) 426 H S R C₂₄H₁₉F₈N₃O₇ 569.5570(M + H) 568(M − H)

[0472] Although the invention has been described with reference to theexamples provided above, it should be understood that variousmodifications can be made without departing from the spirit of theinvention. Accordingly, the invention is limited only by the claims.

We claim:
 1. A compound of the following formula:

wherein: A is a natural or unnatural amino acid of Formula IIa-i:

B is a hydrogen atom, a deuterium atom, alkyl, cycloalkyl, phenyl,substituted phenyl, naphthyl, substituted naphthyl, 2-benzoxazolyl,substituted 2-oxazolyl, (CH₂),cycloalkyl, (CH₂).phenyl,(CH₂)_(n)(substituted phenyl), (CH₂)_(n)(1 or 2-naphthyl),(CH₂)_(n)(substituted 1 or 2-naphthyl), (CH₂)_(n)(heteroaryl),(CH₂)_(n)(substituted heteroaryl), halomethyl, CO₂R¹², CONR¹³R¹⁴,CH₂ZR¹⁵, CH₂OCO(aryl), CH₂OCO(heteroaryl), or CH₂OPO(R¹⁶)R¹⁷, where Z isan oxygen or a sulfur atom, or B is a group of the Formula IIIa-c:

R¹ is alkyl, cycloalkyl, substituted cycloalkyl, (cycloalkyl)alkyl,substituted (cycloalkyl)alkyl, phenyl, substituted phenyl, phenylalkyl,substituted phenylalkyl, naphthyl, substituted naphthyl, (1 or 2naphthyl)alkyl, substituted (1 or 2 naphthyl)alkyl, heterocycle,substituted heterocycle, (heterocycle)alkyl, substituted(heterocycle)alkyl, R^(1a)(R^(1b))N, or R^(1c)O; R¹ is hydrogen, alkyl,phenyl, substituted phenyl, naphthyl, substituted naphthyl, heterocycleor substituted heterocycle; or R¹ and R¹′ taken together with thenitrogen atom to which they are attached form a heterocycle orsubstituted heterocycle; R² is hydrogen, lower alkyl, cycloalkyl,(cycloalkyl)alkyl, phenyl, substituted phenyl, phenylalkyl, substitutedphenylalkyl, naphthyl, substituted naphthyl, (1 or 2 naphthyl)alkyl, orsubstituted (1 or 2 naphthyl)alkyl; and wherein: R^(1a) and R^(1b) areindependently hydrogen, alkyl, cycloalkyl, (cycloalkyl)alkyl, phenyl,substituted phenyl, phenylalkyl, substituted phenylalkyl, naphthyl,substituted naphthyl, (1 or 2 naphthyl)alkyl, substituted (1 or 2naphthyl)alkyl, heteroaryl, substituted heteroaryl, heteroaryl)alkyl, orsubstituted (heteroaryl)alkyl, with the proviso that R^(1a) and R^(1b)cannot both be hydrogen; R^(1c) is alkyl, cycloalkyl, (cycloalkyl)alkyl,phenyl, substituted phenyl, phenylalkyl, substituted phenylalkyl,naphthyl, substituted naphthyl, (1 or 2 naphthyl)alkyl, substituted (1or 2 naphthyl)alkyl, heteroaryl, substituted heteroaryl,(heteroaryl)alkyl, or substituted (heteroaryl)alkyl; R³ is C₁₋₆ loweralkyl, cycloalkyl, phenyl, substituted phenyl, (CH2)_(n)NH₂,(CH₂)_(n)NHCOR⁹, (CH₂)_(n)N(C═NH)NH₂, (CH₂)_(m)CO₂R², (CH₂)_(m)OR¹⁰,(CH₂)_(m)SR¹¹, (CH₂)_(n)cycloalkyl, (CH₂)_(n)phenyl,(CH₂)_(n)(substituted phenyl), (CH₂)_(n)(1 or 2-naphthyl) or(CH₂)_(n)(heteroaryl); R^(3a) is hydrogen or methyl, or R³ and R^(3a)taken together are —(CH₂)_(d)— where d is an interger from 2 to 6; R⁴ isphenyl, substituted phenyl, (CH₂)_(m)phenyl, (CH₂)_(m)(substitutedphenyl), cycloalkyl, or benzofused cycloalkyl; R⁵ is hydrogen, loweralkyl, cycloalkyl, phenyl, substituted phenyl, (CH₂)_(n)cycloalkyl,(CH₂)_(n)phenyl, (CH₂)_(n)(substituted phenyl), or (CH₂)_(n)(1 or2-naphthyl); R⁶ is hydrogen, fluorine, oxo, lower alkyl, cycloalkyl,phenyl, substituted phenyl, naphthyl, (CH₂)_(n)cycloalkyl,(CH₂)_(n)phenyl, (CH₂)_(n)(substituted phenyl), (CH₂)_(n)(1 or2-naphthyl), OR¹⁰, SR¹¹ or NHCOR⁹; R⁷ is hydrogen, oxo (i.e., ═O), loweralkyl, cycloalkyl, phenyl, substituted phenyl, naphthyl,(CH₂)_(n)cycloalkyl, (CH₂)_(n)phenyl, (CH₂)_(n)(substituted phenyl), or(CH₂)_(n)(1 or 2-naphthyl); R⁸ is lower alkyl, cycloalkyl,(CH₂)_(n)cycloalkyl, (CH₂)_(n)phenyl, (CH₂)_(n)(substituted phenyl),(CH₂)_(n)(1 or 2-naphthyl), or COR⁹; R⁹ is hydrogen, lower alkyl,cycloalkyl, phenyl, substituted phenyl, naphthyl, (CH₂)_(n)cycloalkyl,(CH₂)_(n)phenyl, (CH₂)_(n)(substituted phenyl), (CH₂)_(n)(1 or2-naphthyl), OR¹², or NR¹³R¹⁴; R¹⁰ is hydrogen, lower alkyl, cycloalkyl,phenyl, substituted phenyl, naphthyl, (CH₂)_(n)cycloalkyl,(CH₂)_(n)phenyl, (CH₂)_(n)(substituted phenyl), or (CH₂)_(n)(1 or2-naphthyl); R¹¹ is lower alkyl, cycloalkyl, phenyl, substituted phenyl,naphthyl, (CH₂)_(n)cycloalkyl, (CH₂)_(n)phenyl, (CH₂)_(n)(substitutedphenyl), or (CH₂)_(n)(1 or 2-naphthyl); R¹² is lower alkyl, cycloalkyl,(CH₂)_(n)cycloalkyl, (CH₂)_(n)phenyl, (CH₂)_(n)(substituted phenyl), or(CH₂)_(n)(1 or 2-naphthyl); R¹³ is hydrogen, lower alkyl, cycloalkyl,phenyl, substituted phenyl, naphthyl, substituted naphthyl,(CH₂)_(n)cycloalkyl, (CH₂)_(n)phenyl, (CH₂)_(n)(substituted phenyl), or(CH₂)_(n)(1 or 2-naphthyl); R¹⁴ is hydrogen or lower alkyl; or R¹³ andR¹⁴ taken together form a five to seven membered carbocyclic orheterocyclic ring, such as morpholine, or N-substituted pip erazine; R¹⁵is phenyl, substituted phenyl, naphthyl, substituted naphthyl,heteroaryl, (CH₂)_(n)phenyl, (CH₂)_(n)(substituted phenyl), (CH₂)_(n)(1or 2-naphthyl), or (CH₂)_(n)(heteroaryl); R¹⁶ and R¹⁷ are independentlylower alkyl, cycloalkyl, phenyl, substituted phenyl, naphthyl,phenylalkyl, substituted phenylalkyl, or (cycloalkyl)alkyl; R¹⁸ and R¹⁹are independently hydrogen, alkyl, phenyl, substituted phenyl,(CH₂)_(n)phenyl, (CH₂)_(n)(substituted phenyl), or R¹⁸ and R¹⁹ takentogether are —(CH═CH)₂—; R²⁰ is hydrogen, alkyl, phenyl, substitutedphenyl, (CH₂)_(n)phenyl, (CH₂)_(n)(substituted phenyl); R²¹, R²² and R²³are independently hydrogen, or alkyl; X is CH₂, (CH₂)₂, (CH₂)₃, or S; Y¹is O or NR²³; Y² is CH₂, O, or NR²³; a is 0 or 1; b is 1 or 2, providedthat when a is 1 then b is 1; c is 1 or 2, provided that when c is 1then a is 0 and b is 1; m is 1 or 2; and n is 1, 2, 3 or 4; or apharmaceutically acceptable salt thereof.
 2. The compound of claim 1wherein A is


3. The compound of claim 2 wherein R³ is lower alkyl.
 4. The compound ofclaim 2 wherein R^(3a) is hydrogen.
 5. The compound of claim 1 whereinR¹ is phenyl, substituted phenyl, naphthyl, substituted naphthyl,heterocycle, or substituted heterocycle.
 6. The compound of claim 1wherein R¹′ is hydrogen.
 7. The compound of claim 1 wherein R¹′ is loweralkyl.
 8. The compound of claim 1 wherein R¹ and R¹′ taken together withthe nitrogen atom to which they are attached form a heterocycle orsubstituted heterocycle.
 9. The compound of claim 1 wherein B isCH₂O(2,3,5,6-tetrafluorophenol).
 10. The compound of claim 1 wherein Bis hydrogen.
 11. The compound of claim 1 wherein R² is hydrogen.
 12. Thecompound of claim 1 in the cyclic ketal form and having the followingstructure:


13. The compound of claim 12 wherein B is lower alkyl or benzyl.
 14. Apharmaceutical composition comprising a compound of claim 1 incombination with a pharmaceutically acceptable carrier.
 15. A method fortreating an autoimmune disease, comprising administering an effectiveamount of the pharmaceutical composition of claim 14 to a patient inneed thereof.
 16. A method of treating an inflammatory disease,comprising administering an effective amount of the pharmaceuticalcomposition of claim 14 to a patient in need thereof.
 17. A method oftreating a neurodegenerative disease, comprising administering aneffective amount of the pharmaceutical composition of claim 14 to apatient in need thereof.
 18. A method of preventing ischemic injury to apatient suffering from a disease associated with ischemic injury,comprising administering an effective amount of the pharmaceuticalcomposition of claim 14 to a patient in need thereof.
 19. A method forexpanding of hematopoietic cell populations or enhancing their survival,comprising contacting the cells with an effective amount of thepharmaceutical composition of claim
 14. 20. The method of claim 19wherein the cell populations are granulocytes, monocytes, erthrocytes,lymphocytes or platelets for use in cell transfusions.
 21. A method ofprolonging the viability of an organ that has been removed from a donoror isolated cells derived from an organ for the purpose of a futuretransplantation procedure, comprising applying an effective amount ofthe pharmaceutical composition of claim 14 to the organ or isolatedcells to prolong the viability of the same as compared to untreatedorgan or isolated cells.
 22. The method of claim 21 wherein the organ isan intact organ.
 23. The method of claim 21 wherein the isolated cellsare pancreatic islet cells, dopaminergic neurons, blood cells, orhematopoietic cells.